Novel reassortant of H9N2 avian influenza viruses isolated from chickens and quails in Egypt

Research (Published online: 20-08-2021)

21. Novel reassortant of H9N2 avian influenza viruses isolated from chickens and quails in Egypt

Moataz Elsayed, AbdelSatar Arafa, Shahira Abdelwahab, Amro Hashish and Ahmed Youssef

Veterinary World, 14(8): 2142-2149

ABSTRACT

Background and Aim: Poultry infections with H9N2 avian influenza viruses (AIVs) are endemic in Egypt. This study determined the genetic changes in the sequences of H9N2 AIVs isolated from chicken and quails in Egypt, including determining genetic reassortment and detecting the main genetic changes in hemagglutinin (HA) and neuraminidase (NA) genes.

Materials and Methods: Swab samples were collected from chicken and quails, examined through reverse transcription-polymerase chain reaction, and AIVs from positive samples were isolated in embryonated chicken eggs. Complete genome sequencing and phylogenetic analyses were conducted for two H9N2 AIV isolates, and sequences of HA and NA gene segments were analyzed in another two isolates.

Results: A novel reassortant virus was identified from a commercial chicken flock (A/chicken/Egypt/374V/2016) and quails from a live bird market (A/quail/Egypt/1253V/2016). The reassortant viruses acquired four genome segments from the classic Egyptian H9N2 viruses (HA, NA, NP, and M) and four segments from Eurasian AIVs (PB2, PB1, PA, and NS). Many genetic changes have been demonstrated in HA and NA genes. The isolated novel reassortant H9N2 virus from quails showed amino acid mutations in the antigenic sites on the globular head of the mature HA monomer matched with the parent Egyptian H9N2 virus.

Conclusion: This work described the genetic characterization of a novel reassortment of the H9N2 virus in Egypt. The emergence of new reassorted AIV viruses and genome variability raises the concern of an influenza pandemic with zoonotic potentials.

Keywords: genes, H9N2, influenza, mutations, poultry, reassortment.

Discrepancies in the efficacy of H5 inactivated avian influenza vaccines in specific-pathogen-free chickens against challenge with the Egyptian H5N8 clade 2.3.4.4 Group B virus isolated in 2018

Research (Published online: 20-08-2021)

20. Discrepancies in the efficacy of H5 inactivated avian influenza vaccines in specific-pathogen-free chickens against challenge with the Egyptian H5N8 clade 2.3.4.4 Group B virus isolated in 2018

Amena Abd El-Moeid, Ayman Hany EL-Deeb, Marwa Fathy Elsaied, Reem Ahamed Soliman, Mounir Mohamed EL-Safty and Hussein Aly Hussein

Veterinary World, 14(8): 2131-2141

ABSTRACT

Background and Aim: Highly pathogenic avian influenza H5N8 virus of clade 2.3.4.4 was newly emerged to Egypt and firstly detected in carcasses of wild birds in November 2016. This study assessed the protection efficacy and virus shedding reduction of three different inactivated avian influenza (AI) H5 (H5N1, H5N2, and H5N3) commercial vaccines against challenge with two newly emerging highly pathogenic AI virus H5N8 Egyptian isolates in specific-pathogen-free (SPF) chicks.

Materials and Methods: 10-day-old SPF chicks (n=260) were divided into 20 groups (n=13). Groups 1-5 were vaccinated through the subcutaneous route (S/C) with 0.5 mL of H5N1 vaccine, Groups 6-10 were vaccinated (S/C) with 0.5 mL of H5N2 vaccine, and Groups 11-15 were vaccinated (S/C) with 0.5 mL of H5N3 vaccine. Positive control groups (16-19) were challenged at 25 and 31 days old (2 and 3 weeks post-vaccination [PV]) using H5N8 clade 2.3.4.4 A/duck/Egypt/ F13666A/2017(H5N8) and H5N8 clade 2.3.4.4 A/chicken/Egypt/18FL6/2018(H5N8). Group 20 was left non-vaccinated as a control. All vaccinated groups were divided and challenged with both viruses at 25 and 31 days of age. The viral challenge dose was 0.1 mL of 106 EID50/0.1 mL titer/chick, and it was administered oronasally. All chicks were kept in isolators for 14 days after each challenge. Sera samples were collected weekly and at 2 weeks post-challenge (PC) to detect a humoral immune response. PC mortalities were recorded daily for 10 days to calculate the protection percentages. Tracheal swabs were collected from the challenged chicks in different groups at 3, 5, 7, and 10 days PC. Kidneys and spleens were collected at 3, 5, 7, and 10 days PC and kept in formalin for histopathological examination to assess lesions and severity scores. Tracheal swabs were inoculated in 10-day-old SPF embryonated chicken eggs for virus titration and to calculate shedding levels.

Results: All studied vaccines displayed 70-100% protection within 10 days PC. Hemagglutination inhibition results from sera samples revealed antibody titers ranging from 0.6 to 5.4 log2 starting at 1-week PV with the highest titers at 4 weeks PV. Challenged SPF chickens exhibited a notable reduction in virus shedding, with an average of 1.5-2 log10, compared to control birds. Various histopathological lesions with different scores were detected.

Conclusion: Our findings suggest that the inadequate virus shedding reduction and protection efficacy of studied vaccines were variable and that the type of vaccine to be used under field conditions should be reconsidered. Study of the variability between the Egyptian old emerged AI (AIV) 2017 H5N8 strains and the new emerging AIV 2018 H5N8 is required to achieve optimal protection and limit the current economic losses.

Keywords: avian influenza, avian influenza viruses H5N8, clade 2.3.4.4, HPAI, H5 vaccines.

Correlation between age, testosterone and adiponectin concentrations, and sperm abnormalities in Simmental bulls

Research (Published online: 18-08-2021)

19. Correlation between age, testosterone and adiponectin concentrations, and sperm abnormalities in Simmental bulls

Abdullah Baharun, Syahruddin Said, Raden Iis Arifiantini and Ni Wayan K. Karja

Veterinary World, 14(8): 2124-2130

ABSTRACT

Background and Aim: Capacity for sperm production is affected by age, which is related to the morphology of sperm abnormalities and can affect fertility. The aim of this study was to evaluate the relationship between age and concentrations of testosterone and adiponectin with sperm abnormalities in Simmental bulls.

Materials and Methods: The study used 11 bulls, separated into three groups. The first group consisted of five bulls aged 4-5 years, and the second and third groups each consisted of three bulls, aged 6-7 and 8-10 years, respectively. The average sperm motility of the animals ranged from 57.66±2.60% to 70.17±0.22%. Blood samples were obtained from the coccygeal region of the animals. Testosterone and adiponectin analysis was performed using the enzyme-linked immunosorbent assay method. Sperm morphology was evaluated using carbol fuchsin-eosin staining according to the Williams method. Finally, correlations between testosterone and adiponectin concentrations, age, and sperm abnormalities were analyzed using Pearson's correlation analysis.

Results: The findings revealed a significant correlation (p<0.01) between the concentrations of testosterone and adiponectin (–0.538), age (–0.588), and abnormal sperm morphology (–0.912). Moreover, they revealed that the concentration of testosterone in the bulls aged 8-10 years was lower, at 21.89±4.56 ng/mL, compared to that in the bulls aged 4-5 years, at 36.15±1.29 ng/mL, and 6-7 years, at 35.16±5.39 ng/mL. The findings also revealed a positive correlation between adiponectin concentration and age (0.529) and sperm abnormalities (0.506). The increase in testosterone concentration was inversely related to the adiponectin concentration (–0.538). Moreover, the mean amount of abnormal sperm increased with increasing age: 3.82±0.33% in the group aged 4-5 years, and 4.40±0.72% and 10.20±1.97% in the groups aged 6-7 years and 8-10 years, respectively.

Conclusion: The study data indicate that there is a decrease in testosterone concentration, a high adiponectin concentration, and an increase in abnormal sperm with increasing age in bulls.

Keywords: adiponectin, age, Simmental bull, sperm abnormalities, testosterone.

Biochemical and physicochemical indicators of the quality of milk and meat obtained from cows with brucellosis

Research (Published online: 17-08-2021)

18. Biochemical and physicochemical indicators of the quality of milk and meat obtained from cows with brucellosis

Valeriy Alexandrovich Agoltsov, Stepan Yurievich Veselovsky, Olga Mikhailovna Popova, Tatiana Mikhailovna Giro and Nataliya Victorovna Solotova

Veterinary World, 14(8): 2118-2123

ABSTRACT

Background and Aim: Brucellosis is a disease occurring worldwide. Although it is mainly a cattle disease, it is extremely dangerous for humans. Milk and meat can be contaminated with Brucella. The present study aims to examine the biochemical and physicochemical indicators of the quality of milk and meat obtained from cows positively reacting to brucellosis in comparison with healthy animals.

Materials and Methods: Two groups of cattle meat samples (four muscles from different parts of the carcass) were obtained during slaughter at a specialized meat processing plant, and milk samples were examined from healthy animals (10 cows) and from cows positively responding to brucellosis (10 cows). For the milk samples, federal standards (GOST 32915- 2014 and GOST 25179-2014) and an atomic absorption spectrometer "Kvant-Z ETA" were used. To evaluate the chemical composition of the meat, the "Clover" apparatus and a tissue grinder (SM-3) were used.

Results: In the meat of cows that positively responded to brucellosis, compared with that of healthy animals, the amount of dry matter decreased by 1.2 times, amino ammonia nitrogen by 1.01 times, proteins by 1.2 times, fat by 1.28 times, volatile fatty acids by 1.09 times, tryptophan by 1.25 times, oxyproline by 1.14 times, and protein quality indicator by 1.21 times.

Conclusion: Despite the severity of brucellosis in cattle, the biochemical and physicochemical indicators of the quality of milk and meat obtained from the healthy and contaminated animals vary, although insignificantly.

Keywords: biochemical indicators, brucellosis, hydroxyproline, milk acidity, milk density, tryptophan.

Wound healing potential of human umbilical cord mesenchymal stem cell conditioned medium: An in vitro and in vivo study in diabetes-induced rats

Research (Published online: 17-08-2021)

17. Wound healing potential of human umbilical cord mesenchymal stem cell conditioned medium: An in vitro and in vivo study in diabetes-induced rats

Siufui Hendrawan, Yuyus Kusnadi, Christine Ayu Lagonda, Dilafitria Fauza, Jennifer Lheman, Erwin Budi, Brian Saputra Manurung, Hans Ulrich Baer and Sukmawati Tansil Tan

Veterinary World, 14(8): 2109-2117

ABSTRACT

Background and Aim: Human umbilical cord mesenchymal stem cells (hUC-MSCs) and its conditioned medium (CM) promote wound healing. This study investigated the wound healing potential of hUC-MSC CM in vitro and in vivo using diabetic animal models.

Materials and Methods: The CM from hUC-MSC CM prepared under hypoxic conditions (hypoxic hUC-MSC) was evaluated for stimulating rat fibroblast growth, collagen production (in vitro), and wound healing in animal models (in vivo). An excision wound on the dorsal side of the diabetes-induced rats was established, and the rats were randomly divided into non-treatment, antibiotic, and hypoxic hUC-MSC CM groups. The cell number of fibroblasts and collagen secretion was evaluated and compared among the groups in an in vitro study. By contrast, wound size reduction, width of re-epithelialization, and the collagen formation area were assessed and compared among the groups in an in vivo study.

Results: CM under hypoxic conditions contained a higher concentration of wound healing-related growth factors. Hypoxic hUC-MSC CM could facilitate fibroblast cell growth and collagen synthesis, although not significant compared with the control group. Re-epithelialization and collagen production were higher in the hUC-MSC CM group than in the antibiotic and non-treatment groups.

Conclusion: Hypoxic hUC-MSC CM possessed more positive effects on the wound healing process based on re-epithelialization and collagen formation than antibiotic treatment did.

Keywords: conditioned medium, diabetic induced rat, human umbilical cord mesenchymal stem cells, wound healing.

Subclinical mastitis: Prevalence and risk factors in dairy cows in East Java, Indonesia

Research (Published online: 16-08-2021)

16. Subclinical mastitis: Prevalence and risk factors in dairy cows in East Java, Indonesia

Himmatul Khasanah, Hidayat Bambang Setyawan, Roni Yulianto and Desy Cahya Widianingrum

Veterinary World, 14(8): 2102-2108

ABSTRACT

Background and Aim: Subclinical mastitis (SCM) is a disease that frequently attacks lactating dairy cows and possibly decreases production, causing economic losses to farmers. This study aimed to define the prevalence of SCM and risk factor-associated related management practice by dairy farmers in East Java, Indonesia.

Materials and Methods: The milk sample per quarter of individual lactating Friesian–Holstein cows (592 samples) was collected from 148 cows from 62 selected farms in the region with high dairy cattle populations in Malang, Sidoarjo, Mojokerto, Probolinggo, Pasuruan, Lumajang, Jember, and Banyuwangi. SCM determination was performed using the California mastitis test. A survey including field observation and interviews with farmers was conducted to find out the management practices of the selected farms. The analysis of risk factors was conducted by multinomial regression using the IBM SPSS version 26.0 (IBM SPSS Statistics, Chicago, USA).

Results: In addition, 68.18% and 66.72% at the cow and quarter levels, respectively, of the lactating dairy cows examined had SCM. In multinomial regression, four variables were significantly given impact in association with the SCM prevalence in identifying related lactation stage (odds ratio [OR]: 1-2 months=5.67, 2-6 months=9.435), teat wiping after milking (OR=42.197), house cleanliness (OR: dirty=0.120, moderate=0.527), and location (regencies) of raising the cows (OR: Sidoarjo=0.076, Mojokerto=0.165, Jember=1.210, Probolinggo=3.449, Lumajang=1.638, Malang=1.210, and Pasuruan=0.681).

Conclusion: The SCM prevalence in East Java is relatively high and a threat to the dairy industry's performance. This study found a significant association with SCM that needs to be considered in the practice of management to prevent and control SCM. However, the finding also suggested that hygienic management practices performed by farmers need to be improved to reduce SCM incidents.

Keywords: California mastitis test, lactating dairy cows, management practice, subclinical mastitis.

Immunodiagnosis of cattle fascioliasis using a 27 kDa Fasciola gigantica antigen

Research (Published online: 16-08-2021)

15. Immunodiagnosis of cattle fascioliasis using a 27 kDa Fasciola gigantica antigen

Mohamed J. Saadh, Samer A. Tanash, Ammar M. Almaaytah, Issam J. Sa'adeh, Saed M. Aldalaen and Khawla D. Al-Hamaideh

Veterinary World, 14(8): 2097-2101

ABSTRACT

Background and Aim: Diagnosis of fascioliasis depends on clinical symptoms and routine laboratory tests. Recently, antibodies and circulating antigens of Fasciola were used for detecting active infections. Therefore, this study aimed to identify Fasciola gigantica antigens in the sera of infected cattle using Western blotting and enzyme-linked immunosorbent assay (ELISA) for an accurate diagnosis of cattle infected with F. gigantica.

Materials and Methods: Serum samples were obtained from 108, 23, and 19 cattle infected with Fasciola gigantica, Paramphistomum cervi, and Strongylids, respectively, including 57 non-infected cattle that were used as healthy cattle for the study. Western blotting and ELISA were then used to detect circulating Fasciola antigens at 27 kDa.

Results: The target epitope was detected in an F. gigantica adult-worm antigen preparation, excretory/secretory products, and serum from cattle infected with F. gigantica. However, it was absent in sera from P. cervi, Strongylids, and healthy cattle. The purified 27 kDa F. gigantica (FPA-27) antigen was also detected in cattle serum using ELISA with high degrees of sensitivity and specificity (94% and 82%, respectively), and the area under the receiver operating characteristic curve was 0.89 with a highly significant correlation of p<0.0001.

Conclusion: The FPA-27 is proposed to be a promising candidate for the serodiagnosis of fascioliasis in cattle.

Keywords: antigen, Fasciola gigantica, fascioliasis, immunodiagnostics.