Journal of World's Poultry Research

545-555

Authors: Lihui Zhu

Fatty liver hemorrhagic syndrome is more common in laying hens with excess body weight (BW) and in the middle and late phase of egg production. However, no specific biomarkers in chickens can be used to diagnose liver steatosis or liver injury. The present study aimed to assess whether BW can be used to predict fatty liver in aged laying hens. This study also searched for potential plasma FLHS biomarkers. For these purposes, correlation among BW, relative weight of liver and abdominal fat, and plasma markers were analyzed in Hy-line brown laying hens. Furthermore, plasma levels of potential biomarkers were analyzed during the formation of fatty liver. Concentrations of triglycerides and total cholesterol were positively associated with BW in aged laying hens, while liver fat deposition was similar among chickens with different BW, indicating that BW cannot be used as the only criterion to discriminate aged laying hens with liver steatosis. A trend of increasing triglyceride, total cholesterol, fatty acidbinding protein 4 (FABP4), and lipoprotein lipase levels was found as age increased, and they were positively associated with BW indicating that they might be risk markers for FLHS in laying hens. The findings indicated that the plasma level of FABP4 was positively associated with the severity of fatty liver in aged laying hens. All the above results suggested that FABP4 might be a potential diagnostic indicator for FLHS.

565-570

Authors: Setsumi Motsoene Molapo

The objective of the present study was to determine the impact of the feeding levels on the reproductive characteristics of Koekoek chickens. A total of 270 Koekoek chickens were randomly assigned to 4 feeding level treatments in a completely randomized design. The four feeding level treatments were fully fed during the rearing and laying phase (AA), fully fed during the rearing phase and restricted feeding during the laying phase (AR), restricted feeding during the rearing phase and fully fed during the laying phase (RA), and restricted both during the rearing and laying phase (RR). The General Linear Model procedure (SPSS software, version 17) was used to analyze the data set. The pubic bone measurements were 23.6, 25.1, 16.1, and 15.1 mm for chickens that received AA, AR, RA, and RR treatments, respectively, at 18 weeks of age. At 32 weeks of age, chickens given AA and RA treatments had wider pubic bones than chickens given AR and RR treatments. Combined ova and oviduct weights were higher in the fully fed chickens at 18 weeks of age. Koekoek chickens in AA treatment had the highest average egg production. Chickens given AR treatment had lower average egg weights than those given AA, AR, and RR treatments. Chickens treated with AA and AR reached puberty earlier than those that were treated with the RA and RR treatments. The eggs produced by chickens given RR treatment had a higher average hatching percentage. The lowest percentage of hatches was observed in chickens that were fed ad libitum during the rearing phase. In conclusion, the feed restriction only during the rearing phase improved the reproduction performance of Koekoek chickens.

580-586

Authors: Freddy Manyeula

Guinea fowls are increasingly popular in Botswana since they not only provide an alternative to access protein in the form of eggs and meat but also become a good source of income for the peasants. There are different varieties of Guinea fowl in Botswana, including pearl grey, lavender, royal purple, and white. Indeed, there is a need to conduct more studies related to the phenotypic correlations among egg quality traits in different varieties of helmeted guinea fowl found in Botswana. Therefore, the present study was targeted toward the evaluation of both the external and internal quality characteristics of the four different varieties of the domesticated helmeted guinea fowl found in Botswana. In this regard, a total of 150 eggs were collected from the so-called varieties of domesticated helmeted guinea fowl. The egg weight was positively and significantly correlated with egg length, egg width, shell weight, egg surface area, and egg volume; however, the egg weight was negatively correlated with egg shape index. Of the four varieties of domesticated helmeted guinea fowl found in Botswana, the white variety had the strongest correlation coefficients with various external egg quality traits and different internal egg quality characteristics. The lavender variety had the highest correlation coefficients with internal and external egg quality traits. It seems that the selection for higher egg weight as is the case in the current egg grading system can lead to the greatest improvements in other egg quality characteristics in the white and lavender varieties, compared to the pearl grey and royal purple varieties. Therefore, the lavender and white varieties are the potential candidates for the possible selection of layer-type guinea fowl varieties.

605-614

Authors: Waleed S. El-Tahawy

Breeding programs play an important role in increasing the performance of chickens. The poultry industry regards growth and reproduction as the two most economically valued characteristics for providing adequate animal proteins. Genetic variations are the basis of animal breeding. The present study was conducted on three genes, including Prolactin, 3-Hydroxy-3-Methylglutaryl-Coenzyme A Reductase (HMGCR), and Gonadotropin-Releasing Hormone Receptor (GNRHR). DNA was isolated from 48 chickens taken from three strains Lohmann Brown (17), Sinai (24), and Gimmizah (7) for Prolactin, HMGCR, and GNRHR gene amplification by using the PCR protocol. Electrophoresis was performed on the PCR products and the bands were viewed on a transilluminator. The size of the Prolactin gene, HMGCR, and GNRHR were 154, 675, and 210 bp, respectively. For the Sinai strain, five bands for Prolactin, two bands for HMGCR, and six bands for GNRHR were found while for the Lohmann Brown strain, five bands of Prolactin gene, three bands for HMGCR, and five bands for GNRHR were found. Regarding the Gimizah strain, two bands were found for Prolactin and GNRHR genes and there was only one band for the HMGCR gene. The Lohmann Brown strain respectively matured 13 and 91 earlier than Gimizah and Sinai strains with a higher egg number during the first 90 days.

623-630

Authors: Peymaneh Davoodi

Alternative chicken production systems have become popular in recent years due to animal welfare criteria and consumer’s perceptions. General beliefs express that the meat quality of chicken reared under free-range systems is better than that of chickens under conventional production conditions. The aim of this study was to compare the meat quality and carcass traits of chickens raised in conventional and free-range systems. Either conventional or free-range systems used meat-type Hubbard JA57 birds with a slaughter age of approximately 78 days. For assessing carcass traits and meat quality, six male chickens were selected from each system. The meat quality parameters, pH at 45 minutes, ultimate pH, color coordinates, drip loss, cooking loss, and water-holding capacity were measured. Furthermore, proximate parameters, such as crude protein, total fat, and crude ash were determined. There were no significant differences in main carcass yield and breast muscles between chickens reared in two systems, however, color values dramatically were influenced by rearing systems. Breast muscle samples from birds reared under the conventional system had a smaller hue angle and saturation value than those from the free-range birds. Moreover, the drip loss parameter was significantly higher in free-range chickens. The ash and protein contents of breast muscles were similar although raw breast meat from free-range birds had significantly lower fat content. The results prove that a free-range rearing system can modify the appearance, color values, and fat content of chicken meat and it can be a part of the interests of meat production consumers.

643-648

Authors: Dina Novitasari

Avian Influenza (AI) has been spread rapidly in almost all the provinces of Indonesia by the end of 2007, and it has become endemic. Avian Influenza viruses can be infecting to human from direct or indirect contact with the infected or dead poultry, and a visit to the wet poultry market in the neighborhood. Seroprevalence studies can be used to identify the clinical key, epidemiological studies, and the spread of AI viruses in humans. The aim of present study was to investigate the seroprevalence of Avian Influenza (AI) virus, subtype H5 among poultry workers at the central traditional market in industrial. To meet the mentioned demand, 26 blood samples were collected from the poultry workers via the median cubital vein. The antibody titer was examined using Hemagglutination Inhibition (HI) assay using H5 antigen from duck licensed under A/Dk/Indonesia/AU-78/12 (H5N1) and three kinds of red blood cells taken from horse, chicken and guinea pig. The serum samples were added with Receptor Destroying Enzyme (RDE) with a ratio of 3:1 (v/v) for an overnight, and pretreated with 10% of red blood cells before the HI assay was conducted. Based on the findings, it can be concluded that the percentage of seroprevalence of Avian Influenza (AI) virus, subtype H5 among poultry workers at central traditional market was 0%. Thirteen samples showed a negative result of the HI test. All of the workers’ blood serum obtained less than 24 antibody titer from the HI test. As the results showed, the research on the poultry workers in the traditional market was carried out, and it obtained negative results; all the workers were not infected with the Avian Influenza virus. In other words, Avian Influenza is not meaningful in poultry farm workers in Sidoarjo suburb traditional farms.

407-428

Authors: Liao Y

Much like other indigenous domesticated animals, Guangxi Three-yellow chickens (GX-TYC) in China have experienced strong selective pressure, and show specific phenotypic changes in physiology, morphology and behavior. To identify genomic footprints or selection signatures left by artificial selection during domestication of GX-TYC, the whole genomes of 12 GX-TYC hens were sequenced to executed selective sweep analyses and gene functional enrichment analysis (Gene Ontology and Kyoto Encyclopedia of Genes and Genome pathways). A total of 10.13 million single nucleotide polymorphisms and 842,236 insertion/deletion polymorphisms (Indels) were found. Forty-six windows showed a Z score of heterozygosity (ZHp) lower than -5, which potentially were considered to be positively selected regions. Gene annotation identified 55 genes in these regions. Selection signatures were found mainly on the SSC5, SSC8, SSC23 and SSCZ. GO and KEGG analyses revealed that these genes were related to growth, immune responses as well as carbohydrate, lipid and amino acid metabolisms. In addition, two genes, fructose-1,6-bisphosphatase 1 and fructose-1,6-bisphosphatase 2 were enriched into four signaling pathways, three of which are involved in carbohydrate metabolism and insulin signaling. SHC3, FANCC and PTCH1, in combination with FB1 and FBP2, were clustered together in a region of chromosome Z, and thus might have been selected together. The results have uncovered some genetic footprints of chicken domestication, providing not only an important resource for further improvements of fowl breeding, but also a useful framework for future studies on the genetics of domestic chickens as well as on the phenotypic variations and certain diseases of chickens.

436-442

Authors: Ardyansyah RH

The current study aimed to evaluate the effect of Agaricus bisporus and Auricularia auricula crude extracts as feed additives on the growth performance of broiler chickens. The samples included 240 one-day-old chicks randomly divided into 8 dietary treatments, each treatment consisted of 3 replicates with 10 chicks per replicate. The dietary treatment groups were control group (T0), basal diet + zinc bacitracin (T1), basal diets + 0.4% Agaricus bisporus extract (T2), basal diet + 0.8% Agaricus bisporus extract (T3), basal diet + 1.2% Agaricus bisporus extract (T4), basal diet + 0.4% Auricularia auricula extract (T5), basal diet + 0.8% Auricularia auricula extract (T6), and basal diet + 1.2% Auricularia auricula extract (T7). The measured variables included feed intake, body weight gain, feed conversion ratio, and production index. In addition, the study aimed to evaluate the reducing sugars level, antioxidant IC50, and antimicrobial efficacy of mushroom extracts prepared using three different solvents (i.e., water, ethanol, and methanol). The findings indicated that methanolic extract contained higher reducing sugars and had better antimicrobial efficacy. The results of experimental research revealed that mushrooms crude extracts had no significant effects on the growth performance of broiler chickens.

451-455

Authors: Pancapalaga W

The present study aimed to examine the effects of fermented soy isoflavones on the poultry feed towards the quality of eggs. A total of 100 Isa Brown chickens aged 32 weeks were divided into 4 groups and 5 replicates, including T0 (control feed without the provision of fermented soy isoflavones), T1 (feed with 4% of fermented soy isoflavones), T2 (feed with 8% of fermented soy isoflavones), and T3 (feed with 12% of fermented soy isoflavones). The treatments were given for 10 weeks. The observed chemical qualities of eggs included HDL, LDL, isoflavones in egg yolks, physical quality (e.g., their weight and eggshell thickness). All the data were analyzed by using analysis of variance. The results showed that the administration of fermented soy isoflavones in poultry feed had no significant effect on egg weight and eggshell thickness (p> 0.05), but had a significant effect on HDL, LDL, and isoflavones in egg yolk (p< 0.01). Based on the results, it can be concluded that 80 mg/100 g of the fermented soy isoflavones (equivalent to the addition of 12%) in laying hens’ feed would increase the content of isoflavones and egg yolks’ HDL, reduce egg yolks’ LDL, and lead to no significant change in the weight and thickness of eggshells.

469-479

Authors: Syed B

In recent years, probiotics and synbiotics have gained considerable interest in poultry feeding as an alternative to antibiotics due to antibiotic resistance concerns. The objective of this dual study was to evaluate the efficacy of synbiotic supplementation alone or in combination with different Antibiotic Growth Promoters (AGPs), compared to the untreated control group of broiler chickens production performance. In the first experiment, a total of 1260 one-day-old male Ross 308 broiler chickens were randomly assigned to 7 diet treatments, with 6 replicates per diet treatment and 30 birds per replicate over a 42-day period. The diet treatments included a control diet based on corn-soybean without additives (T1), and the diet treatment with bacitracin (BMD 100 ppm, T2), colistin (10 ppm, T3), synbiotic (PoultryStar me, 0.5 kg/t, T4), a combination of synbiotic (0.5 kg/t) and bacitracin (60 ppm, T5), synbiotic (0.5 kg/t) and colistin (5 ppm, T6), synbiotic (0.5 kg/t), bacitracin (60 ppm), and colistin (5 ppm, T7). During the critical period of rearing from hatch to day 10, the synbiotic supplementation resulted in a significantly higher body weight gain than its combination with bacitracin. No other dietary treatment showed a remarkable improvement in the body weight gain, feed intake, or feed conversion ratio, compared to the only synbiotic application (T4) during the entire trial period. The tendency towards an improved feed conversion ratio was observed during the use of symbiotic (T4, 1.87), compared to the control group (T1, 1.93) during the entire trial period. Compared with the control group (T1, 2.78%), broiler mortality was also lower in the synbiotic group (T4, 1.11%). In the second experiment, a total of 1500 one-day-old male Ross 308 broiler chickens were randomly assigned to 4 diet treatments; with 15 replicates per diet treatment, and 25 birds per replicate over a 42-day period. The dietary treatments included a control group diet based on corn-soybean without additives (T1), and the treatment diets with bacitracin (BMD 1000 ppm, T2), synbiotic (PoultryStar me, 0.5 kg/t, T3), and a combination of synbiotic (0.5 kg/t) plus bacitracin (BMD 1000 ppm T4). Birds fed antibiotic or synbiotic alone or in a combination had numerically a higher body weight and an average daily gain than the control group. There was a tendency of improvement in the feed conversion ratio during the age of 1-24 days, and throughout the experimental period. The evaluated synbiotic could serve as an effective alternative to AGPs, such as bacitracin and colistin in broiler chicken diets, especially during the first crucial period. The synbiotic can serve this purpose without combining it with AGPs, such as colistin or bacitracin.

485-492

Authors: El-Kholy KH

The objective of this study was to determine the optimal level of coconut oil (CO) supplementation in the diet to enhance the performance of Domyati ducks. A total number of 300 Domyati ducks (240 females and 60 males) aged 25-week-old were randomly assigned to 4 experimental groups of three replicates, each replicate included 5 males and 20 females of Domyati ducks. The groups received CO at 0, 1.0, 1.5, and 2.0% during the experimental period. The results indicated that egg weight, egg number, and egg mass significantly increased in treated groups, compared to the control group. Furthermore, fertility and hatchability percentages were superior in 1.0, and 1.5% CO groups, compared to other experimental groups. Low-density lipoprotein in ducks that received CO was significantly lower than that of the control group. It is concluded that the inclusion of CO at a 1.5% level could be enough and useful for improving the reproductive and physiological performance of Domyati ducks.

507-512

Authors: Castillo-Grijalva M

Avian Influenza and Newcastle disease are the two most important diseases of poultry and are globally considered as threats to public health and economy. There is little information published about these diseases in peacocks and other common backyard poultry in Guatemala. Therefore, an exploratory serosurvey was conducted to determine the presence of circulating antibodies to Avian Influenza (AI) and Newcastle Disease (ND) viruses in a semi-captive population of peacocks in southwestern Guatemala. Additionally, the circulation of antibodies to these pathogens in backyard chickens, ducks, and turkeys from a neighboring community was explored. Blood samples were obtained from 48 peacocks, 30 chickens, 6 ducks, and 4 turkeys. The samples were processed in the Regional Reference Laboratory for Animal Health, at the Veterinary Medicine Faculty, University of San Carlos of Guatemala, located in Guatemala City. Antibodies against AI virus were investigated by Agar Gel Immunodiffusion, and antibodies against ND virus were examined using Hemagglutination Inhibition. No antibodies against AI virus were detected. Most of the samples (97.7%) were negative for antibodies against ND virus, except for two turkeys that carried low antibody titers. The findings of the present study indicate that no virulent strains of AI or ND viruses were circulating in the investigated site.

520-526

Authors: Mohammed MH

Surveillance studies for Newcastle disease virus (NDV) are critical to monitor the potential spreading of these viruses among wild birds as well as domestic poultry. This study was conducted to determine the incidence of NDV in wild birds in Egypt in 2016. Out of 159 collected samples from eight different species of wild birds, six (3.77%) samples were positive for paramyxoviruses by semi-nested RT-PCR assay based on the RNA-dependent RNA polymerase gene. Of six positive samples, four NDVs were successfully isolated in 11-day-old specific-pathogen-free embryonated hens’ eggs. Partial sequences of the fusion gene of the four isolates were amplified using RT-PCR. Phylogenetic analysis of partial sequences of RNA-dependent RNA polymerase gene and fusion genes indicated that the detected NDV viruses in wild birds in Egypt are related to class I NDVs strains. Four Egyptian NDV isolates from wild birds exhibited sequence motif of 111GERQER↓LVG119 at the cleavage site as lentogenic virus in wild birds. Continuous active surveillance may help better monitoring of NDVs circulating in wild birds before newly emerging viruses in domestic poultry.

125-139

Authors: Rabie TSKM.

The present study was intended to estimate the specific genetic variants by using nine genetic markers among five rabbit breeds (New Zealand White, California, Chinchilla, Flander, and Babion) in Egypt. A total of 128 animals were used (19-35 rabbits per breed). A total of 97 alleles were detected across the breeds and the average number of alleles per locus was 2.16±0.11. Five private alleles were present in Babion breed, where the locus INRACCDDV0023 had two private alleles of 293 and 297 base pairs with allele frequencies of 0.4 and 0.1, respectively. The INRACCDDV0036, INRACCDDV0304, and INRACCDDV0241 loci had private allele for each (185bp (freq: 0.24), 197 (freq: 0.47), and 137bp (freq: 0.26), respectively). The mean of He values ranged from 0.35±0.06 to 0.49±0.07. The average of the polymorphic information content was 0.41 (ranged from 0.298 at INRACCDDV0211 to 0.599 at INRACCDDV0036 locus). To estimate the genetic deviation of the five rabbit breeds, two parameters were evaluated: genetic differentiation (FST), and genetic distance. The FST values varied from 0.029 (INRACCDDV0036) to 0.785 (INRACCDDV0022). The similarity matrix showed that the Chinchilla breed was distinct from other breeds. In addition, among the nine loci, the Hardy-Weinberg equilibrium was highly significant for five loci. Therefore, the rabbit breeds are good reservoirs of allelic diversity that is the major basis for genetic improvement. Consequently, the breeders need a formal conservation plan for such breeds that are in danger of extinction in near future.

145-150

Authors: Berghiche A

To evaluate the impact of uncontrolled use of veterinary drugs on broilers in eastern Algeria, an experimental plan was developed for the evaluation and identification of drug toxicity in 60 chickens (30 treated and 30 non-treated with antibiotics) using analysis of serum biochemical parameters, autopsy, morpho-metric and histopathological analysis of certain internal organs. The results of the serum biochemical analysis revealed that the uric acid and aspartate aminotransferase values in antibiotic-treated chickens were high, while the lesion status showed a dominance of respiratory lesions, followed by digestive lesions, particularly hepatic lesions. The morphometric study of the internal organs (liver, kidney, and intestine) demonstrated that abnormal liver appearance was very important with minor atrophic changes in the kidney, while the histopathological examination of the liver revealed the presence of deposition in the center of the hexagons in the apical area with an apparent homogeneous structure of fibrous connective tissue. Also, there were apparent deep sinus defects in peripheral areas with an overload of fibrin. The histopathological examination of the kidneys revealed proximal tubular atrophy in the renal parenchyma along with loss of distal intratubular consistency to the peripheral zone of homogeneous structure persuading the peripheral edema. It is concluded that the uncontrolled use of antibiotics in the poultry industry leading to a moderate to severe toxicity.

165-171

Authors: Mohamed FM

The aim of the present work was to determine antibacterial sensitivity and resistance patterns of Pasteurella multocida isolated from rabbits in different farms of Assiut Governorate. Also, this study aimed to detect virulence-associated gene (toxA) of Pasteurella multocida. A total of 40 freshly dead rabbits were used to collect samples from liver, lung and subcutaneous abscess. In addition, tracheal swab samples were collected from 20 diseased rabbits. Bacteriological examination revealed that Pasteurella spp. were isolated and phenotypically identified with an incidence rate of 55% (33 out of 60 rabbits). Ten Pasteurella spp. isolates were randomly chosen for antibiotic sensitivity testing and molecular identification using PCR. Antibiotic sensitivity test was carried using standard disk diffusion method against 13 antibacterial drugs to determine antibacterial sensitivity and resistance patterns of Pasteurella isolates and revealed variable sensitivity and resistance to antibacterial drugs. Pasteurella multocida isolates were sensitive to wide variety of antibiotics (norfloxacin, enrofloxacin, ciprofloxacin, florfenicol, doxycycline, gentamycin, cephradine and cefoxitin). Three out of ten isolates were molecularly confirmed to be Pasteurella multocida and all of them demonstrated the presence of toxA virulence genes. In conclusion, the prevalence of Pasteurella infections in rabbits in Assiut Governorate was relatively high.

180-183

Authors: Abdulla NR

The present study was conducted to determine the effect of DL- and L-methionine on growth performance, carcass characteristics, and gut morphology during the finisher phase in the tropical environment. A total of 560 one-day-old broiler chicks (Cobb 500) were purchased and raised for 35 days. The chicks were divided into four dietary treatments with seven replicates (20 birds per replicate). The basal diet was offered to the chickens during the starter and finisher phases. The DL-methionine was supplemented to the finisher diet as at 0.260% (T1) and 0.179% (T2). Correspondingly, the L-methionine was supplemented to the finisher diet with the same ratios; 0.260% (T3) and 0.179% (T4). The findings revealed no significant differences in growth performance between the two forms of methionine. The obtained results indicated no significant differences in carcass characteristics, the villi heights and crypt depth among the dietary treatments. In conclusion, DL-methionine can be used in broiler nutrition as substitute for L- methionine which is more expensive in poultry industry.

195-202

Authors: Shakal M

Newcastle Disease Virus (NDV) is still a major concern for the Egyptian poultry industry in spite of the mass vaccination programs implemented from a long years ago. The current study aimed to carry out the molecular identification of surface glycoprotein genes of NDV field strain isolated from the Giza governorate, Egypt. Tracheae were collected from 10 broilers NDV-vaccinated chicken flocks (at least three samples from each flock) suffering from mild to moderate respiratory symptoms; with mortalities varying from 10-40% during October 2019. Only five samples showed HA positive activity after propagation in specific pathogen-free embryonated chicken eggs and only one sample was positive for Avian avulavirus 1 by real-time reverse transcription-PCR. Sequencing for the cleavage site of the F protein gene of the positive isolate showed the typical known sequence of velogenic NDV strains (112RRQKRF117). Phylogenetic analysis of both F and HN genes showed high similarity and close relation to Chinese strains of Genotype VII and more specifically subtype VIId, suggesting the role of migratory wild birds in NDV evolution in Egypt. In conclusion, further epidemiological and surveillance studies are strongly recommended to define the exact role of migratory wild birds in NDV evolution in Egypt.

214-222

Authors: Ali NM

Salmonella enterica is considered a zoonotic pathogen that acquires antibiotic resistance in livestock. In the current study, a total of 18 Salmonella enterica isolates recovered from cloacal swabs of diseased and freshly dead broilers were serotyped and assessed for susceptibility to clinically important antibiotics. The multi-resistant isolates were examined for the presence of the antiseptic resistance genes including quaternary ammonium (qacEΔ1) and class 1 integron-integrase (intI1) by PCR. The results of serotyping of 18 Salmonella isolates indicated that five isolates belonged to Salmonella Typhimurium, four isolates belonged to each of Salmonella Kentucky and Salmonella Enteritidis, three isolates belonged to Salmonella Molade and one isolate belonged to each of Salmonella Inganda and Salmonella Larochelle. Fifteen Salmonella isolates (83.3%) were multi-resistant to at least three antibiotics with a multidrug resistance index value of 0.473. All of the intI1-positive strains carried qacEΔ1, confirming that the qacEΔ1 gene is linked to the integrons. The study concluded that the presence of the qacEΔ1 resistance gene and class 1 integrase in multi-drug resistant Salmonella strains might be contributed to co-resistance or cross-resistance mechanisms.

235-246

Authors: Elnabarawy AM

This research was designed to evaluate the effect of naturally contaminated feed with mycotoxins on the efficacy of vaccination against coccidiosis in broilers. Two hundred day-old Hubbard broiler chicks were divided into four groups (50 chicks/group). Groups 1 and 3 were kept on naturally contaminated diets containing 4 ppb aflatoxin, 3 ppb ochratoxin, 1 ppm zearalenone and 2 ppb aflatoxin, 6 ppb ochratoxin and 1 ppm zearalenone in starter and grower feed, respectively. Groups 2 and 4 were fed on diet without detectable levels of mycotoxins. Group 1 and 2 were vaccinated with anticoccidial vaccine at 4 days of age. All groups were challenged with Eimeria tenella (5×104/chick) 14 days post-vaccination. Vaccinated mycotoxicated birds showed a significant reduction in body weight, high mortality, significant oocysts shedding, severe hemorrhagic typhlitis, marked lymphoid depletion in bursa of Fabricius and degenerative changes in liver and kidney. In addition, a remarkable decrease in length and width of intestinal villi, mucosal length and crypt depth. Feed contamination with multi-mycotoxins in permissible level caused vaccination failure and a remarkable decrease in intestinal morphometric histopathological parameters.

250-262

Authors: Abou Zeid MAM

The present study aimed to investigate the incidence of Salmonella infection in diarrheic ducklings in Kafr El Sheikh Governorate, Egypt. A total of 100 samples were collected from ducklings suffered from diarrhea and mortality. Also, 50 litter samples were collected from duck farms. All specimens were collected under aseptic conditions for the isolation of Salmonella spp. The incidence of Salmonella was 7% in pooled samples from cecum, liver, spleen and gall bladder and 6% in litter samples. Ten strains of Salmonella spp. were serotyped, of which, S. Salamae (1 strain), S. Miami (2 strains), S. Kentucky (4 strains), S. Paratyphi (2 strain) and S. Magherafelt (1 strain) were detected. Susceptibility of Salmonella isolates to 10 antimicrobial agents showed that Salmonella isolates were highly sensitive to amikacin (100%), followed by trimethoprim/sulphamethoxazole and gentamicin (50%). While isolates showed the highest percentage of resistance to norfloxacin (90%), followed by ciprocin (70%), flumox (70%) and amoxicillin-clavulanic acid (70%). Virulence genes (invA, hilA, and fimA) were detected by PCR assay, all 10 Salmonella isolates showed positive results for three virulence genes, which gave specific amplicon at 284, 150, and 85 base pairs, respectively. Lethality test in five groups of three-day-old ducklings with different five isolated strains indicated a mortality rate ranged from 20-30 % in three isolates only. The most lethal strain S. Paratyphi A was chosen for further investigation as a pathogenicity test. IL-6 slightly decreased in the infected group in comparison to control. The results indicated that ducks infected with Salmonella spp. significantly showed lower RBCs, Hb, PCV, Phagocytic activity, phagocytic index, and serum albumin while, significantly had higher WBCs, neutrophil, lymphocyte, serum globulin, uric acid, creatinine, AST and ALT concentrations compared to non-infected. It could be concluded that Salmonella has hepatic and renal destructive effects and immunosuppressive effects.

278-284

Authors: Shakal M

Antibiotic-resistant bacteria have become one of the major issues and concerns worldwide. For the past years, scientists have investigated the use of treatments in the nano-scale. Nanomaterials, such as metal oxide nanoparticles, have shown promising results due to their antibacterial properties. The aim of this study was to investigate the efficiency of in vitro antibacterial activity of zinc oxide nanoparticles (ZnO NPs) alone and in combination with different antibiotics against avian pathogenic Escherichia coli. In this study, ZnO NPs were synthesized using direct precipitation method. Physical characteristics of ZnO NPs were confirmed using X-ray diffraction and transmission electron microscopy. Antibacterial resistance pattern of 10 antibiotics including amoxicillin, ciprofloxacin, enrofloxacin, gentamicin, doxycycline, levofloxacin, trimethoprim/sulfamethoxazole, tetracycline, spiramycin, and streptomycin, in addition to different concentrations of ZnO NPs, was determined by disc diffusion method on 10 avian pathogenic E. coli (APEC). The results showed that 50% of the strains were resistant to all antibiotics, while the rest were found to be sensitive to one or two antibiotics. The best concentration of ZnO NPs was 50 mg/disk, which showed greater zones than that of other used concentrations (25, 12.5, 6.25, 3.125, and 1.56 mg/disk). The combination of spiramycin and gentamycin with ZnO NPs showed a synergistic effect while the combination of ZnO NPs with ciprofloxacin, enrofloxacin, and streptomycin showed an antagonistic effect. No antibacterial effect was observed in combination of ZnO NPs with other used antibiotics. This study recommends in vivo evaluations to confirm the results.

292-298

Authors: Abd El Tawab AA

Bacillus cereus is an opportunistic pathogen that can cause food poisoning in humans as a result of consuming foods containing toxins or bacteria. In this study, the incidence of B. cereus and its virulence genes in meat products was investigated. Isolation of B. cereus was performed using selective PEMBA media and confirmed by morphological and biochemical tests and Vitek2 compact system. The incidence of B. cereus strains in beef and chicken meat products was 28%. The incidence of Bacillus cereus in frozen rice kofta, frozen kobiba-shami, chicken pane, and chicken nuggets was 16%, 24%, 28%, and 44%, respectively. Moreover, the result of multiplex PCR of virulence genes of groEL gene (533bp), Hbl gene (1091 bp), Nhe gene (766 bp) and Cytk gene (421bp) indicated that groEl gene, Nhe gene, Cytck gene was found in 100% of B. cereus isolated from different meat products, while Hbl gene was detected in 10% of isolates. The results demonstrate that meat products represent a threat to public health through the transmission of Bacillus cereus.

326-335

Authors: Yu Guo, Shuying Huo

In order to reveal the functions of Epimedium (EPI) on the recovery of the reproductive system of laying hens, 120 Hy-Line Brown laying hens (43-week-old) were forced molt through feed withdrawal for 14 days. After the molting period, layer hens were divided into four groups: Dark-control group in a dark environment without any treatment; dark-EPI group in a dark environment and with EPI treatment; light-control group without any treatment under normal illumination (16L: 8D); light- EPI group with EPI treatment under normal illumination (16L: 8D). EPI treatment was maintained for 15 days and each hen received 2 ml of EPI extract (1 g/mL) daily. The rate of egg production was calculated every day. At the end of the experiment, estrogen receptor alpha mRNA, estrogen receptor beta mRNA, and progesterone receptor mRNA in the albumen secreting part and uterus of the oviduct were detected by q-PCR, and the level of serum progesterone, estrogen and luteinizing hormone was measured by enzyme-linked immunosorbent assay. The results showed that EPI effectively improved the laying rate of hens both in dark groups and light groups by promoting the recovery of the oviduct and follicle maturation. In addition, EPI promoted the secretion of estrogen and progesterone both in dark and light groups and improved the expression of estrogen receptor alpha and progesterone receptor in the light group. The results of the experiment provide a good reference for using EPI to improve the development and recovery of the reproductive system of layer hens

342-347

Authors: Omidiwura BRO

In a 42-day feeding trial, the effect of L-Dopa on the performance, serum cholesterol and intestinal microbial load in laying hens were investigated. One hundred and twenty layers aged 34 weeks old were allocated to five dietary treatments with eight replicates and three birds per each replicate. The birds were fed diets supplemented with graded levels of L-Dopa (0, 0.1, 0.2, 0.3 and 0.4%) in a completely randomized design. Performance parameters were monitored. On the day 42, egg, meat and blood samples were obtained to determine total cholesterol and lipoproteins using standard procedures. The ileal digesta was collected for microbial analysis. The results indicated that inclusion of L-Dopa in the diet did not affect the performance parameters and egg cholesterol profile. Serum cholesterol levels of birds fed the control diet and those on 0.1% and 0.2% L-Dopa supplemented diets were similar, but significantly higher than those fed 0.3% and 0.4% L-Dopa. The levels of cholesterol and low-density lipoproteins in the meat of the layers fed on the control diet were significantly higher than those on 0.1% and 0.3% L-Dopa, but were similar to the birds on other treatment diets. The highest and the lowest population of Escherichia coli were found in the birds on the control diet and 0.3% L-Dopa supplemented diet, respectively. The layer hens on L-Dopa supplemented diets had a significantly higher Lactobacillus count than those on the control diet. In conclusion, 0.3% L-Dopa inclusion considerably improved the cholesterol profile in the blood and meat, reduced the population of E. coli, and effectively increased the population of Lactobacillus in the laying hens.

359-366

Authors: Dukhnytskyi V

In this article an attempt was made to present the results of toxicity assessment of insecticides from the group of neonicotinoids, Mospilan RP (active substance acetamiprid) and Actara 25 WG (active substance thiamethoxam) in white mice. The aim was to investigate the acute toxicity of Mospilan RP and Actara 25 WG in white mice. The half-lethal dose (LD50) of Mospilan RP was found to be131.25 ± 34.12 mg/kg Body Weight (BW) per active substance and 656.25 ± 170.6 mg/kg BW per drug, and the Lethal Dose (LD100) was 200 mg/kg BW by the active ingredient (1000 mg/kg BW by drug). The half-life dose (DL50) of Actara 25 WG for white mice was 907.81 ± 24.03 mg/kg BW for the active substance, and 3631.24 ± 96.12 mg/kg BW for the drug; The lethal dose (LD100) was also 1200 mg/kg BW per active ingredient (4800 mg/kg BW per drug). The acute course of poisoning by Mospilan RP and Actara 25 WG in mice was characterized mainly by nervous disorders (lesions of the central and peripheral nervous systems), which is evidenced by the clinical features of poisoning including depression, convulsions, ataxia (impaired movement coordination), tremor and impaired breathing. It has been established that Mospilan RP belongs to the third toxicity class according to the Hygienic classification of substances by skin-resorptive toxicity (DL50> 2000 mg/kg).

371-377

Authors: Nuraini N

The aims of present study was to determine the effect of dietary supplementation of palm oil waste fermented with Phanerochaete chrysosporium and Neurospora crassa (POWF) with ratio 1:1 on performance and some egg characterizes of laying Japanese quails. This experiment used 200 quails (Coturnix-coturnix japonica) aged 20 weeks, with 70% egg production. This study used an experimental method with a completely randomized design (CRD) with four treatments (0%, 8%, 16%, and 24% POWF and five replications). The results of this research showed that the use of POWF in the diet was not affected to feed consumption, daily egg production, egg mass, egg weight, and feed conversion. But, a significant difference in blood total cholesterol, LDL, HDL, egg cholesterol, and index egg yolk color was obsereved, and no significant effect on triglycerides and egg yolk fat. In conclusion, feeding the mixture of palm oil waste fermented with Phanerochaete chrysosporium, and Neurospora crassa with ratio1:1 can be used up to 24% in laying quail rations, without any adverse effect on egg production. It may decrease the blood cholesterol and egg yolk cholesterol.

385-390

Authors: Sahan Yapicier O

Gallibacterium anatis causes infections in the reproductive tract of egg-laying hens and it is associated with increased mortality and decreased egg production. For this study we used singeleplex and multiplex PCR with specific primers to assess the presence of tetracycline resistance (Tcr) (tet A, B, C, D, E, G, H, K, L, M, O, S, P, Q and X), virulence [cytotoxic (RTX-like toxin, gtxA) and fimbrial (flfA)] genes and antibiotic resistance in G. anatis isolates. Among the 20 isolates tested, the highest antimicrobial resistance patterns were observed in erythromycin, streptomycin, tilmicosin (100%) followed by colistin sulphate (65%), cephalexin and tulathromycin (50%). Among 20 isolates examined, 10 (50%) carried tetracycline resistance genes, 7 (35%) had tet(B), 2 (10%) had tet(G), and 1 (5%) had tet(A), (D), (M) or (L). Of these G .anatis isolates were carried out 6 (30%) gtxA but none of flfA gene. Based on present results, it is concluded that virulence and Tcr genes could contribute to pathogencity of G. anatis, which is a major risk to poultry health.

397-406

Authors: Santos TTdos

An experiment was designed to determine the immune profile of broiler chickens between hatch and 9 days of age when fed diets with different fibre and betaine concentrations. A total of 240 day-old Cobb 500 male chickens were allocated to 16 cages with 15 chickens each. Treatments were arranged in a 2 x 4 factorial design, with 2 replicate cages per treatment. Treatments consisted of two feed formulations (low and high fibre diets) and four levels of betaine (0, 1, 3 or 5 kg/tonne). Before the start of trial (hatch), 10 broilers were weighed and blood samples were collected by cardiac puncture, then euthanised by cervical dislocation and jejunal samples collected for the determination of gene expression of claudin 1, claudin 5, occludin and interleukin 2 by PCR. Mononuclear cells populations in the blood samples were determined by flow cytometry. On days 4 and 9, five birds/cage (10 birds/treatment) were selected, euthanised and samples taken as per the start of hatch. Gene expression of claudin 1, claudin 5 and occludin was reduced between 4 and 9 days, independent of the group tested, while interleukin tended to increase between hatch and 4 days and decreased thereafter. Betaine inclusion reduced claudin 5 and occludin gene expression and increased CD8- CD28+ and CD8+ CD28+, suggesting it may aid in accelerating maturation of both the gastrointestinal tract and immune system for broilers in the early days post-hatch.

01-11

Authors: Liu Y, Zhao J, Wang F, Zhou J, Yang X and Yang X

Folic acid had been reported to develop much metabolic regulation function in animals and human beings due to its roles in one carbon metabolism. The current study was conducted to explore folic acid regulation function in primary chicken hepatocytes via supplement and deprivation culture models based on proteomic analysis. Results have shown that folic acid supplement significantly increased intracellular folic acid, 5-Me-THF and SAM contents when compared with folic acid free group (P < 0.05). Whereas, there was no difference about genome 5mC levels and DNMTs mRNA expression between these two groups. Proteomic analysis found 85 differential expressed proteins with 35 down and 50 up regulation. COG and KEGG pathway analysis revealed that amino acid metabolism, carbohydrate metabolism and antioxidant function were affected by folic acid. Posttranslational modification, protein turnover, chaperones and transcription were gathered by COG analysis in relative high proportion. PRMT7 and ARID4B which were associated with histone methylation were up-regulated in the folic acid supplement group, suggesting that folic acid was likely to take part in metabolism regulation of hepatocytes via histone methylation manner in the study. In conclusion, proteomic analysis found 85 differential expressed proteins in hepatocytes with folic acid free and supplementation medium. Folic acid might be involved in amino acid and carbohydrate metabolism and oxidation resistance by its epigenetic modifications functions. Our study also provided fundamental differential protein profiles mediated by folic acid, which can facilitate the understanding of folic acid regulation function in hepatic metabolism.

17-27

Authors: Rasheed AR, Al-Ajeeli KS and Al-Azawy AKh.

A local isolate of low pathogenic Avian Influenza Virus (AIV) H9N2 subtype was used in experimental infection of 50 domestic and 50 feral pigeons (Columba livia domestica) to determine the susceptibility of these birds to H9N2 infections and to study its histopathological effects on vaccinated and unvaccinated pigeons with H9N2 commercial vaccine. The birds were divided into five groups. Groups A and C contained 20 feral pigeons, B and D contained 20 domesticated pigeons. Group E contained 10 feral and 10 domesticated pigeons that were used as unvaccinated controls. Groups A and B were vaccinated with H9N2 and Newcastle disease virus commercial vaccines. Group C and D were vaccinated with Newcastle disease virus vaccine only. All groups except E were challenged with a local isolate of H9N2 serotype. Antibodies titers against AIV were estimated pre and post-vaccination using ELISA. The results indicated low antibody titers against AIV in all groups in pre-vaccination that ranged between 152.83 ± 42.01 and 337.00 ± 150.76 with no significant differences between groups. Post-vaccination antibody evaluation indicated high titers of anti-AIV antibodies in groups A and B (740.13 ± 214.38 and 673.00 ± 242.40, respectively) in comparison to pre-vaccination levels. Clinical signs appeared on 5th day post-vaccination that included mild respiratory signs, digestive disorders, and conjunctivitis in some birds of all groups. Histopathological changes in affected tissues appeared as moderate to severe multifocal necrosis diffused in the parenchymal cells of lung tissues. Infiltration with mononuclear inflammatory cells was detected in some lung tissue areas. Necrotic foci and mononuclear cell infiltration were also observed in trachea and liver of infected pigeons but mild changes were observed in intestine. The challenge virus was re-isolated in embryonated hen's eggs of nine days old by inoculation in allantoic cavity using samples collected from tissues and cloaca of infected pigeons showing clear clinical signs. The re-isolated virus was detected by the haemagglutination test using chicken RBCs and identified by haemagglutination inhibition test using a locally prepared hyperimmune serum to H9N2 in rabbits. In conclusion, pigeons are susceptible to AIV (H9N2) that might facilitate the transmission of the virus to other bird species.

36-40

Authors: Erwan E, Adelina T, Koto A and Maslami V.

L-citrulline (L-Cit) is considered one of the potential amino acids that are able to reduce body temperature in layer chicks. However, there are no reports on functions of L-Cit in KUB chicks. Therefore, the present study aimed to evaluate the effects of oral administration of L-Cit on feed intake, body temperature and plasma biochemical parameters of KUB chicks over two hours post-administration. Neonatal KUB chicks were orally administered with different doses of L-Cit (0, 3.75, 7.5 and 15 mmol/kg body weight). At 30, 60 and 120 min, feed intake was determined, also rectal temperature of chicks was measured by using a digital thermometer. At 120 min after administration, the blood samples were immediately collected through the jugular vein, then biochemical parameters of plasma (total cholesterol, triacylglycerol and total glucose) were analyzed. The obtained results indicated that feed intake, body temperature, and plasma metabolites were not significantly influenced by different doses of L-Cit. In conclusion, L-Cit could not able to improve feed intake, decrease body temperature and change plasma metabolites in KUB chicks.

52-56

Authors: Al-Shadeedi ShMJ.

The present study was carried out to compare the physicochemical characteristics of eggs of guinea fowl, turkeys and domestic chickens outdoor reared in traditional farms in Baghdad, Iraq. A total of 166 fresh eggs; 32 eggs from guinea fowls (Numida meleagris), 44 eggs from turkeys (Meleagris gallopavo) and 90 eggs from domestic chickens; were collected. Egg weight, percentage of egg components, chemical composition (protein, lipids, and ash), and lipid profile were determined. Results revealed the significant differences in egg weight among studied birds. The average egg weights for guinea fowl, turkey, and indigenous chicken were 83.15 ± 0.72, 92.41 ± 0.78 and 61.28 ± 0.62 g, respectively. No significant differences were found in egg components and the chemical composition of the edible portions of the eggs among studied birds. However, the lipid profile of egg yolk indicated that egg cholesterol and LDL levels were significantly higher in guinea fowl and turkey compared with those in indigenous chickens, whereas native chicken has high values of HDL compared to guinea fowl and turkey. There were no significant differences in the triglyceride level in egg yolks among the studied fowls. In conclusion, although egg weight was significantly different among studied birds, eggs of guinea fowl, turkeys, and domestic chickens were similar in nutritional components.

63-70

Authors: Al-Ajeeli KS, Al-Azawy AKh and Ghazuan H.

Chicken Anemia Virus (CAV) infects many bird species worldwide and causes immunosuppression. This condition can facilitate the infection of affected birds with other pathogens including bacteria, viruses, and fungi. No data were available on detection or isolation of CAV from birds in Iraq, therefore this study was designed to detect CAV antibodies in broilers and layers in some poultry farms. Accordingly, 200 samples were collected from broiler and layer farms (100 samples each) from different districts of Diyala province and subjected to the ELISA test. Also, 50 tissue samples from embryonated eggs from different hatcheries, four commercial viral vaccines, and 30 ELISA positive samples were subjected to PCR assay to detect the CAV DNA. The results showed that all of broiler and layer farms sampled were serologically positive for CAV antibodies. The overall seropositivity for CAV antibodies for both chicken breeds was 51.5%. In broilers, 43 out of 100 serum samples were positive for CAV antibodies, whereas 60 out of 100 serum samples from layers were CAV antibody-positive. According to age groups, significant differences were observed among one-week-old broilers (30.2%) compared to other age groups. In layers, the age group of 30 weeks showed a seropositivity rate of 33.3%. Conventional PCR test indicated that all tissue samples collected from suspected birds and embryonated eggs were negative for CAV DNA, but only 2 out of 30 serum samples were PCR positive. It is concluded that CAV is endemic in poultry farms of Iraq and may facilitate the vaccination failure against other viruses.

81-86

Authors: Hilmi M, Prastujati UA, Khusnah A, Khirzin MH and Yannuarista D.

Cheese production waste contains carbon source, one of which is lactose as an energy source in the development of fermentation, especially in the manufacture of probiotics for poultry. The research method used 3% fermented cheese whey in 17-week-old laying hen drinking water to analyze the productivity of laying hens and microbiology. The purpose of the research is to determine the effective concentration of fermented whey cheese to improve productivity, physical and chemical composition of eggs, and evaluate fat metabolism in laying hens. The present study used 120 laying hens aged 17 weeks. The experimental method was designed using the T-test method of control (P0), fermented whey cheese (P1). The observed variables were productivity (feed consumption, water consumption, daily egg production, feed conversion ration), and microbiology of small intestine and excreta (lactate acid bacteria, salmonella, Escherichia coli.). The effect of adding fermented whey cheese into drinking water decreased feed consumption and FCR in contrast to the control group (P0) but increased egg production, egg weight, and egg mass weight. The effect of adding fermented whey cheese into drinking water decreased the number of, Escherichia coli bacteria and increases lactic acid bacteria in the digestive tract of broiler chickens, especially in the ileum, caeca, so also with those in the stool. The number of Salmonella Bacteria was significantly decreased and very significantly increased lactic acid bacteria in the feces during the fermentation of whey cheese. The decrease in ammonia in the stool was very significant compared to the control group. In conclusion, adding fermented whey cheese into drinking water can reduce feed consumption, feed conversion ratio (FCR), Escherichia coli, salmonella, ammonia (NH3), and increase egg production, egg weight, egg mass weight, final body weight, and lactate acid bacteria. The use of fermented whey cheese can be used as a nutraceutical feed additive to inhibit pathogenic bacteria in the intestine and increase lactic acid bacteria.

96-101

Authors: Nuraini N, Shafan Nur Y and Djulardi A

Improving the quality of cocoa pods through fermentation with Pleurotus ostreatus reduced its crude fiber content, especially for lignin and cellulose. Fermentation of cocoa pods product increased their utilization in poultry diet. An experiment was conducted to determine the effect of feeding cocoa pods waste fermented by Pleurotus ostreatus on the laying performance and egg quality of quail. This experiment employed completely randomized design with five dietary treatments: 0%, 5%, 10%, 15% and 20% cocoa pods fermented by Pleurotus ostreatus in the diets and four replications. 200 laying quail (7 weeks of age) were randomly allocated into 5 treatments (10 birds per treatment) and 4 replications. Diets included iso nitrogen 20% and iso metabolism energy 2800 kcal/kg. The results of the experiment indicated that feed intake, hen-day egg production, egg weight, egg mass production, and feed conversion were not affected but egg cholesterol was affected by increasing cocoa pods fermented products in the diet. In this experiment, a diet which utilized 15% cocoa pods fermented using Pleurotus ostreatus maintained the laying performance of quail and reduced egg yolk cholesterol (20.30%).

109-117

Authors: Mousa MR, Mohammed FF, Abdel reheem FA, El-deeb AH and Ahmed KA

Velogenic Newcastle disease virus (VNDV) shows systemic dissemination and it affects the histological structure and function of the kidneys. Present study is an novel attempt to correlate the changes in renal biochemical functions and gene expression of different inflammatory cytokines with related renal pathology induced by VNDV in experimentally infected broilers in different ages. One hundred broiler chickens were divided into 4 groups: group 1 served as control (non-infected group), and groups 2, 3 and 4 were inoculated with 106 EID50 of NDV on the 10th, 20th and 30th days of age, respectively. Serum samples were collected to evaluate uric acid and creatinine levels. Kidney specimens were collected for virus detection using real time RT-PCR, evaluation of gene expression of IL-6 and IFN-γ as well as histopathological, histochemical and immunohictochemical analysis. The highest concentration of uric acid and creatinine profile were found in Group 4 (12.06 ± 1.25 and 2.16 ± 0.12, respectively) on 7 dpi. IL-6 and IFN-γ were significantly elevated at the 7th dpi in Group 4 compared to other infected groups. The renal histopathological lesions included tubulointerstitial nephritis and glomerulopathy with expression of virus antigen in different areas of renal tissue. histopathology was the hallmark of VNDV infection in broiler chickens; their severity was related to the increased expression of inflammatory cytokines genes (IL-6 and IFN-γ) and virus antigen residence in renal tissue.