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Porcine circovirus type 2 and its associated diseases in southwestern Nigeria: Farmers' perception and level of awareness
2022
Oluwawemimo Oluseun Adebowale (Department of Veterinary Public Health and Preventive Medicine, Federal University of Agriculture, Abeokuta, Nigeria) | Olufemi Samuel Amoo (Centre for Human Virology and Genomics Research, Nigerian Institute of Medical Research NIMR, Yaba, Nigeria) | Kayode Olayinka Afolabi (Molecular Epidemiology and Public Health Research Group (MEPHREG), Department of Biological Sciences, Anchor University, Nigeria. & Anchor University Center for Global Health (AUCGH), Nigeria) | Abimbola Adetokunbo Oloye (Department of Veterinary Surgery and Theriogenology, Federal University Agriculture, Abeokuta, Nigeria.)
Objective: Porcine circovirus type 2 (PCV2) is one of the most important causative agents of swine diseases that pose a global economic threat. Presently, there is little or no information on the perception and awareness of PCV2 and its associated effects among pig farmers in Nigeria. Therefore, this research was carried out to describe pig farmers' views, awareness, and likely impact of PCV2 and its associated postweaning multisystemic wasting syndrome (PMWS) on pig production in the southwestern region of Nigeria. Materials and Methods: A cross-sectional survey of pig farmers in Oyo and Ogun states, Southwest Nigeria, was carried out with the help of a self-administered questionnaire. Results: A total of 111 farms out of the 385 required took part in the study, resulting in a total response rate of 28.8%. 89 (79.2%, 95% CI = 70.8–85.8) pig farmers who participated were unaware of PCV2, while 46 (41.4%, 95% CI = 32.7–50.7) had heard about PMWS. The level of awareness was generally poor, with an average score of 1.43 (SD ± 1.25; 23.9%). Only 23% (25/111) of the participants had a high level of awareness. To promote awareness about PCV2/PMWS, participants' most preferred sources of information were seminars, extension services (especially by veterinary and agricultural extension officers), social media (WhatsApp and YouTube), and mobile telephone (through calls or text messages). Conclusions: The present study showed a gap in the level of farmers' awareness about PCV2/ PMWS, and to bridge the gap, more scientific-based evidence is needed to promote targeted educational programs and policy formulations. Also, with the dearth of information about PCV2, it is necessary to determine its prevalence and the characteristics of the virus possibly circulating within the swine herds in Nigeria. [J Adv Vet Anim Res 2022; 9(2.000): 203-210]
Show more [+] Less [-]Production performance of Alabio ducks (Anas platyrhynchos Borneo) under different levels of drinking water salinity
2022
Abrani Sulaiman (Department of Animal Science, Faculty of Agriculture, Lambung Mangkurat University, Banjarbaru, Indonesia) | Surya Rahmatullah (Department of Animal Science, Faculty of Agriculture, Lambung Mangkurat University, Banjarbaru, Indonesia) | Hefni Effendi (Center for Environmental Science PPLH, IPB University, Bogor, Indonesia) | Gamaliel Simanungkalit (School of Environmental and Rural Science, University of New England, Armidale, Australia.)
Objective: To examine the effects of the salinity level of drinking water on the egg production and quality of Alabio ducks. Materials and Methods: A total of 60 female Alabio ducks, aged 6 months, were subjected to this study. All ducks were kept in stage-type cages (1 m length × 1 m width × 0.5 m height), where each cage was inhabited by 4 ducks for 56 days of experimentation. All ducks were offered a mixed ration ad libitum for laying ducks, according to the nutritional requirements for egg-type ducks. The treatment in this study was drinking water with five stratified salinity levels, namely P0 = freshwater (0% salinity); P1 = water with a salinity of 0.75 practical salinity unit (PSU) (equal to 0.75 g NaCl/l); P2 = water with a salinity of 1.5 PSU (1.5 gm/l); P3 = water with a salinity of 2.5 PSU (2.5 gm/l); and P4 = water with a salinity of 3 PSU (3.0 gm/l). Observations were made on water intake, feed intake, egg production, and egg quality (egg weight, egg shape index density, shell proportion, shell thickness, yolk index, albumen index, and Haugh unit). Results: The results showed that the difference in salinity levels in drinking water from 0.75 PSU to 3 PSU did not affect water intake, feed intake, egg production, or egg quality of Alabio ducks for the first 56 days of the laying period (p > 0.05). Conclusions: It was concluded that Alabio ducks have a good tolerance for drinking water salinity of up to 3 PSU, or equal to 3 gm/l NaCl. [J Adv Vet Anim Res 2022; 9(2.000): 241-245]
Show more [+] Less [-]Traditional farmers' pig trait preferences and awareness levels toward reproductive biotechnology application in Zambia
2022
Rubaijaniza Abigaba (Department of Animal Science, University of Zambia, Lusaka, Zambia) | Pharaoh Collins Sianangama (Department of Animal Science, University of Zambia, Lusaka, Zambia) | Progress H. Nyanga (Department of Geography and Environmental Studies, University of Zambia, Lusaka, Zambia) | Wilson N. M. Mwenya (Department of Animal Science, University of Zambia, Lusaka, Zambia) | Edwell S. Mwaanga (Department of Biomedical Sciences, University of Zambia, Lusaka, Zambia.)
Objective: This study assessed traditional farmers' preferences for indigenous pigs and their awareness levels toward reproductive biotechnology. Materials and Methods: This cross-sectional, descriptive study employed a mixed-methods con¬current triangulation design with a pragmatic approach. For quantitative data collection, a ran¬domly selected sample size of 622 respondents was interviewed using a structured questionnaire. A semi-structured interview guide was used in seven focus group discussions (FGDs) for qualitative data. For quantitative data, descriptive statistics were used to find out how often something happened, and chi-square tests were used to look for relationships. For qualitative data, thematic analysis was used. Results: The majority (66.9%) of respondents were female, and they had largely (64.8%) attended a primary level of education. A slight majority (43.1%) of the respondents were 30–45 years old. Most respondents (65.1%) were low-income earners (below K500). Incidentally, the majority (74.1%) of respondents had low awareness of reproductive biotechnologies. Respondents' aware¬ness levels were associated with gender (p < 0.001), education (p < 0.001), income (p < 0.01), and not age (p > 0.05). With regard to trait preference, a total of seven indigenous pig traits were reportedly preferred, of which disease resistance (25.9%) and litter size (23.8%) were the most preferred. From FGDs, participants valued indigenous pigs, citing many preferred traits, of which disease resistance and litter size were the most emphasized traits. Some participants were aware of many reproductive biotechnologies and their perceived advantages. However, they were more familiar with artificial insemination, heat detection methods, and synchronization. Conclusions: The majority of respondents had low awareness of reproductive biotechnologies. Respondents' awareness was associated with sociodemographic characteristics. The identified indigenous pig traits preferred by traditional farmers included adaptive and productive traits. [J Adv Vet Anim Res 2022; 9(2.000): 255-266]
Show more [+] Less [-]Evaluation of the in vitro and in vivo inhibitory effects of Artemisia herba-alba against the growth of piroplasm parasites
2022
Rasha Eltaysh (National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan. & Department of Pharmacology, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt) | Mohamed Abdo Rizk (National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan. & Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt) | Shimaa Abd El-Salam El-Sayed (National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan. & Department of Biochemistry and Chemistry of Nutrition, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt) | Khaled Abouelnasr (Department of Surgery, Anesthesiology and Radiology, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt) | Abdelnaser Ahmed Abdallah (Veterinary Teaching Hospital, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt) | Ikuo Igarashi (National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan.)
Objective: The effect of Artemisia herba-alba methanolic extract monotherapy and combination therapies on the in vitro growth of several Babesia and Theileria parasites in vitro and mice was investigated in this study. Materials and Methods: Fluorescence assay using SYBR Green I stain was used to evaluate the antibabesial efficacy inhibitory of A. herba-alba either in vitro or in vivo. Hematological parameters in the treated mice were analyzed using a Celltac MEK-6450 computerized hematology analyzer. Results: Artemisia herba-alba reduced the growth of Babesia bovis, Babesia bigemina, Babesia divergens, Theileria equi, and Babesia caballi in vitro in a dose-dependent manner. The in vitro inhibitory impact of A. herba-alba on B. divergens and B. caballi cultures was amplified when combined with either diminazene aceturate (DA). In B. microti-infected mice, a combination therapy consisting of A. herba-alba and a low DA dose inhibited B. microti growth significantly (p < 0.05) better than treatment with 25 mg kg−1 DA. Conclusions: These data show that A. herba-alba, when paired with a modest DA dose, could be a promising medicinal plant for babesiosis treatment. [J Adv Vet Anim Res 2022; 9(2.000): 267-274]
Show more [+] Less [-]Molecular detection of pathogenic Leptospira spp. in urban rodents from wet markets in northeast Malaysia
2022
Intan Noor Aina Kamaruzaman (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Muhamad Aiman Mohd Mokhtar (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Hong Wei Ting (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Yong Kai Yuan (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Azim Wafiy Gulam Shah (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Tan Wan Loong (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Nurshahirah Shaharulnizim (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Mohd Farhan Hanif Reduan (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Fathin Faahimaah Abdul Hamid (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Nur Amalina Noralidin (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Nur Athirah Abdul Manaf (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Che Wan Salma Che Wan Zalati (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia) | Loong Shih-Keng (Tropical Infectious Diseases Research & Education Centre, Universiti Malaya, Kuala Lumpur, Malaysia) | Simon Clegg (School of Life Sciences, University of Lincoln, Brayford Pool Lincoln, United Kingdom) | Luqman Abu-Bakar (Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu, Malaysia.)
Objective: This short study describes the occurrence of pathogenic Leptospira spp. in two major wet markets in Kota Bharu, Kelantan, Malaysia. Materials and Methods: 30 rodents (20 rats and 10 shrews) were caught in 2 wet markets, and a postmortem was performed to extract both kidneys. Molecular diagnosis via polymerase chain reaction (PCR) was conducted to detect leptospiral DNA using universal and pathogenic Leptospira primers, respectively. Results: The results showed that 20/28 (72%) rat samples were detected positive for Leptospira spp, and all shrews were negative. Further sequencing analysis identified L. interrogans and L. borgpetersenii as the most frequently Leptospirosis species from kidney samples. Conclusions: The presented study here sheds light on the presence of pathogenic leptospires har¬boring the rat population in both wet markets in Kelantan, which presents a great public health risk to wet market workers and visitors. [J Adv Vet Anim Res 2022; 9(2.000): 275-281]
Show more [+] Less [-]Fowl typhoid live lyophilized vaccine applied at 3-month intervals protected layer chickens from Salmonella gallinarum infection and prevented cloacal shedding
2022
Taslima Akter (Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh) | Mohammed Nooruzzaman (Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh) | Sheikh Mohammad Shariful Hoque Belal (Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh) | Mustak Ahammed (Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh) | ABM Jalal Uddin (Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh) | Rokshana Parvin (Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh) | Md. Abu Hadi Noor Ali Khan (Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh) | Md. Ariful Islam (Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh) | Md. Mokbul Hossain (Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh.)
Objective: Here, we developed and tested the efficacy of a vaccination protocol based on a commercially available live attenuated Salmonella enterica serovar Gallinarum (Salmonella galli¬narum) in layer chickens. Materials and Methods: 50 layer chickens of 16 weeks age were obtained and divided into two groups (n = 25), control and vaccinated. The vaccinated group received Salmonella vaccine at 0.2 ml/bird, s/c route at 16, 18, 30, and 42 weeks of age. At 21 weeks of age, birds from both groups were challenged with S. gallinarum orally at 4 × 107 colony-forming unit per bird. Results: Both rapid serum plate agglutination and enzyme-linked immunosorbent assay demonstrated a rising rate of seroconversion in vaccinated birds across the study period, with a 4% positive rate at 18 weeks, 56% at 21 weeks, 60% at 30 weeks, and 64% at each time point of 42 and 54 weeks. The vaccine showed 100% clinical protection and reduced the Salmonella shedding in the feces and eggs of the challenged birds. On the contrary, the unvaccinated challenged birds showed clinical signs and lesions typical of Salmonella infections with morbidity and mortality rates of 36% and 20%, respectively, and had high rates of Salmonella shedding in feces and eggs. Conclusions: With the proposed vaccination schedule Salmonella shedding was prevented, and a high seroconversion was confirmed. To prevent Salmonella infections in laying flocks, a 3-month interval immunization program is advised starting at the pre-laying stage. [J Adv Vet Anim Res 2022; 9(2.000): 301-309]
Show more [+] Less [-]Valorization of cricket, Acheta domesticus (Linnaeus, 1758), flour as a source of dietary protein in Japanese quail, Coturnix japonica (Temminck and Schlegel, 1849), farming
2022
Francois Djitie Kouatcho (Department of Science and Technology of Organic Agriculture, Faculty of Sciences / School of Chemical Engineering and Mineral Industries, University of Ngaoundere, Ngaoundere, Cameroon) | Razvan Mihail Radu Rusu (Department of Management of Animal Productions, Faculty of Food and Animal Sciences, Iaşi University of Life Sciences, Iaşi, Romania) | Bachirou Mohamadou (Department of Biological Sciences, Faculty of Sciences, University of Ngaoundere, Ngaoundere, Cameroon) | Bobga Aoudou (Department of Biological Sciences, Faculty of Sciences, University of Ngaoundere, Ngaoundere, Cameroon) | Ioan Mircea Pop (Department of Management of Animal Productions, Faculty of Food and Animal Sciences, Iaşi University of Life Sciences, Iaşi, Romania) | Marius Giorgi Usturoi (Department of Management of Animal Productions, Faculty of Food and Animal Sciences, Iaşi University of Life Sciences, Iaşi, Romania) | Leonard S. Ngamo Tinkeu (Department of Biological Sciences, Faculty of Sciences, University of Ngaoundere, Ngaoundere, Cameroon.)
Objective: Quail production is ranked as an important alternative animal protein source in Cameroon. One of the main constraints of this production is the quality of feed, which lacks pro¬tein that is regularly supplied by fish meal. To avoid disagreements due to the constant shortage of fish meal, alternative protein sources are needed, and among them are crickets (Acheta domes¬ticus). The goal of this study was to find out how well Japanese quails could be raised if fish meal was replaced with cricket meal. Materials and Methods: A total of 192, 4-week-old quails were divided into 12 similar sets of 8 females and 8 males. The Four experimental diets (T00, T15, T30, and T45) were all formulated based on the level (0%, 15%, 30%, and 45%, respectively) of fish meal substitution with cricket meal in the basal diet (crude protein: 20.18% and ME: 3,013.78 kcal/kg) and randomly assigned to three sets in a completely randomized design consisting of four treatments each repeated three times. Growth, carcass, and some reproduction parameters were collected. The data were analyzed using one-way analysis of variance and the Duncan test, with a significance level of 5%. Results: Cricket meal diets increased body weight in males (204.32 ± 5.69 gm for T45) and regard¬less of the sex (226.72 ± 29.45 gm for T30) compared to 184.17 ± 3.11 gm and 214.55 ± 32.77 gm for the control group, respectively. In females, substitution at 30% increased body weight (253.80 ± 6.48 gm), while 45% induced a reduction (216.67 ± 6.49 gm) when compared to the control group value (244.92 ± 6.07 gm). Carcass yield and the proportion of different parts were not significantly affected by the experimental diets. Liver proportions were significantly higher at 15% and 35% cricket meal incorporation compared to 45%. First songs and egg laying were recorded at 7 weeks with T15, which also led to improved egg laying performance compared to the other treatments. Ovaries were poorly developed in the T45 females compared to the other treatments. Conclusions: Cricket flour might be a good candidate as a locally available protein source to sub¬stitute fish meal in the Japanese quail's diet at the finisher and reproductive stages, and the level of 30% seems to be more efficient. [J Adv Vet Anim Res 2022; 9(2.000): 310-322]
Show more [+] Less [-]Anticoccidial effects of Tinospora rumphii extract in naturally infected goats
2022
Pearl Muyco Saladino (College of Veterinary Medicine, University of Southern Mindanao, Kabacan, Cotabato, Philippines) | Elsa Amarille Gonzaga (Department of Veterinary Paraclinical Sciences, College of Veterinary Medicine, University of Southern Mindanao, Kabacan, Cotabato, Philippines) | Loveille Jun Amarille Gonzaga (Department of Chemistry, College of Science and Mathematics, University of Southern Mindanao, Kabacan, Cotabato, Philippines.)
Objective: This study aimed to determine the anticoccidial potential of Tinospora rumphii ethan¬olic stem extract, resulting in an effective reduction in oocysts per gram counts of Eimeria species in goats in comparison to the commercial coccidiostat toltrazuril. Materials and Methods: Eighteen goats that were naturally infected with coccidia were identified as experimental animals. The experimental animals were grouped and were administered a single dose of T. rumphii extract at concentrations of 300, 600, 1,000, and 2,000 mg extract/kg body weight. The fecal samples were collected before treatment, during the first treatment, and every week thereafter for 28 days posttreatment. Fecal examination was carried out using a modified McMaster technique to determine the oocysts per gram of feces, and the mean efficacies of the extracts were calculated. Results: Stem extracts of T. rumphii were able to lower the oocysts per gram count than pre¬treatment for all concentrations, but the efficacy, in general, was still lower than toltrazuril. A significant difference existed in the efficacy of the extracts among the treatments on day 14. Administering 1,000 mg/kg extract resulted in the highest efficacy rate (95.25%) at 14 days post¬treatment and was comparable to that of toltrazuril (89.89%). Conclusion: The results show that the T. rumphii stem extract has the potential to reduce the Eimeria species oocysts per gram counts of naturally infected goats. [J Adv Vet Anim Res 2022; 9(1.000): 8-12]
Show more [+] Less [-]Acaricide resistance of Rhipicephalus decoloratus ticks collected from communal grazing cattle in South Africa
2022
Mandla Yawa (Department of Livestock and Pasture Science, University of Fort Hare, Alice, South Africa) | Nkululeko Nyangiwe (Dohne Agricultural Development Institute, Stutterheim, South Africa, & Department of Agriculture and Animal Health, University of South Africa, Roodepoort Johannesburg 1710, South Africa) | Ishmael Festus Jaja (Department of Livestock and Pasture Science, University of Fort Hare, Alice, South Africa) | Munyaradzi Christopher Marufu (Department of Veterinary Tropical Diseases, University of Pretoria, Pretoria, South Africa) | Charles T. Kadzere (Dohne Agricultural Development Institute, Stutterheim, South Africa.)
Objective: This study aimed to determine acaricide resistance in Rhipicephalus decoloratus ticks collected from grazing cattle between November 2018 and May 2019 in Elundini, Senqu, and Walter Sisulu Local Municipalities in the northeastern region of the Eastern Cape Province. Materials and Methods: A sample of 20–30 adult engorged female R. decoloratus ticks were collected from at least 10 randomly selected cattle (highly tick-infested) at each dip tank and placed into the labelled plastic collection bottles containing absorbent paper and with a perforated lid at a constant room temperature of ±28°C and >70% relative humidity until resistance testing commenced. The Shaw larval immersion test method was used to determine R. decoloratus larvae resistance to various acaricide concentration levels [amidines, organophosphate (OP), and synthetic pyrethroids (SPs)]. Results: This study found that most ticks were susceptible to exposure to different acaricide field concentrations of amidines (49% at 250 ppm), OPs (33% and 47% at 300 ppm and 500 ppm, respectively), and SPs (44% and 23% at 150 ppm and 300 ppm, respectively). The resistance test¬ing results showed no resistance to amidines at any localities and no resistance to OP in the Senqu region. However, resistance development of the larvae to amines, OPs, and SPs was extensively observed in Senqu (18%, 6%, and 7%), Elundini (15%, 15%, and 17%), and Walter Sisulu (13%, 19%, and 9%) regions, respectively. Conclusions: The larvae's resistance is a cause for worry. Hence, the continuous monitoring of tick resistance to commonly used acaricides will help mitigate widespread acaricidal resistance and sustain livestock productivity. [J Adv Vet Anim Res 2022; 9(1.000): 33-41]
Show more [+] Less [-]Expressions of IL4, IL10, and IFNγ cytokines genes during bacterial mastitis
2022
Rana A. Faaz (Department of Microbiology, College of Veterinary Medicine, University of Basrah, Basrah, Iraq) | Fawziah A. Abdullah (Department of Microbiology, College of Veterinary Medicine, University of Basrah, Basrah, Iraq.)
Objective: Many bacteria are involved in causing mastitis in dairy cows. Perfect identification of bacteria is crucial for the appropriate choice of drug for treatment. This study aims to find out the various bacteria that cause mastitis through the 16S ribosomal ribonucleic acid (16S rRNA) gene. Materials and Methods: A total of 150 mastitis somatic cell samples were tested with bacterial nested polymerase chain reaction (PCR) universal primers, targeting the 16S rRNA gene. The primers had both Gram-positive and Gram-negative bacterial specificities. Inflammatory cytokine interleukin (IL-10), IL-4, and interferon-gamma (IFNγ) expression genes were measured and compared in mastitis-free and mastitis-affected animals. Results: Based on the PCR, 70 (46.7%) samples showed positive results. The expression of the IL-10 gene was significantly higher (p < 0.001) in mastitis-affected cows than noninfected animals. Compared to cows diagnosed with clinical mastitis, the IL-4 and IFNγ genes were expressed more strongly in healthy cows (p < 0.0001). Conclusion: Mastitis has been linked to both Gram-positive and Gram-negative bacteria. These genes are strong predictors of mastitis in the states analyzed, as evidenced by the differential expression in mastitis and healthy conditions of the IL-4, IL-10, and IFNγ genes. The genes examined here and others will be the subject of additional research. [J Adv Vet Anim Res 2022; 9(1.000): 42-52]
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