Introduction: Avian pathogenic Escherichia coli (APEC) is a leading cause of extraintestinal infection and heavy economic losses. Imparting immunity after vaccination with live attenuated strain vaccination is an ideal strategy for infection control. This study considers an FtsK knockout mutant strain as a candidate. Material and Methods: An FtsK knockout mutant of APEC strain E058 was constructed and the pathogenicity of the mutant and wild-type strains was further evaluated in chickens. Results: The 50% lethal doses of each strain for one-day-old specific-pathogen-free (SPF) chickens challenged experimentally via trachea were 10⁵.⁵ and 10⁷.⁰ colony-forming units (CFU) respectively. Chickens challenged with the wild-type strain exhibited typical signs and lesions of avian colibacillosis, while those inoculated with the mutant strain showed mild pericarditis and pulmonary congestion. The growth rate of the FtsK mutant strain was much slower than the wild-type strain in the heart, spleen, liver, and lung of infected chickens. Conclusion: These results indicated that the APEC FtsK mutant can be attenuated for chickens, and that this mutant has the potential for the development of an APEC vaccine.

Akabane virus (AKAV) has been detected in a variety of host species in China, but there are only limited records of its occurrence in goats. However, more attention needs to be paid to understanding the diversity of viruses in this species. The aim of the study was to explore the genotype characteristics and variation trend of AKAV and their relationship with virulence in Yunnan, China. Blood samples were collected from goats during routine surveillance of goat diseases in Yunnan province in 2019. The AKAV CX-01 strain was isolated using BHK-21 cells. To understand pathogenicity, the virus was intraperitoneally (IP) and intracerebrally (IC) inoculated into suckling mice and tissue samples were subsequently analysed histopathologically and immunohistochemically. Akabane virus CX-01 strain induced encephalitis and impairment of the central nervous system with fatal consequences. Phylogenetic analysis based on the ORF sequences of the small segments indicated that the AKAV isolate used was most closely related to the GD18134/2018 Chinese midge and bovine NM BS/1strains, while phylogenetic analysis based on the medium segments showed a close relationship between CX-01 and the Chinese GLXCH01 strain. The CX-01 isolate was related to AKAV genogroup Ia and probably originated from a recombination of different strains.

The pseudorabies virus (PRV) gene encoding thymidine kinase (tk) is an important virulence-associated factor. Attenuation of PRV in susceptible animals is a frequent result of tk deletion. The aim of the study was to assess the pathogenicity of tk-deleted PRV in rats. Sprague Dawley rats were infected with the tk-deleted PRV strain SuHV-1 ΔTK:247via intranasal or intramuscular inoculation. PRV loads in ten tissues from dead and euthanised rats were determined using real-time PCR. Infection with SuHV-1 ΔTK:247 could cause death in rats. The 50% lethal dose (LD₅₀) of SuHV-1 ΔTK:247 via intranasal inoculation was 10³.¹⁶ TCID₅₀ in rats. Intramuscular inoculation required a higher dose of SuHV-1 ΔTK:247 (10⁵.⁰ TCID₅₀). A high SuHV-1 ΔTK:247 titre was observed in the trigeminal ganglia or spinal cord of dead rats. The results of this study show that rats are highly susceptible to PRV infection, and tk deletion did not completely diminish the pathogenicity of PRV in rats.

Classical swine fever virus (CSFV) causes an economically important and highly contagious disease of pigs, leading to economic losses around the world. Attenuated live vaccines with CSFV antigens have played an important role in the prevention and control of the disease. Porcine kidney 15 (PK15) cells have been widely used for the propagation of CSFV, but this cell line is not efficient or homogeneously susceptible to viral infection. To achieve a homogeneous PK15 cell line which enabled high titre replication of CSFV, we used the limiting dilution cell cloning method. We developed two cell clones, PK15-1A6 and PK15-3B1, which respectively have high- and low-permissive phenotypes to CSFV infection. The PK15-1A6, PK15-3B1, and PK15 parent cells showed different characteristics in cell proliferation rate, susceptibility to CSFV infection, and CSFV production. The mean virus titres per millilitre reflected by TCID₅₀ values in PK15-1A6, PK15-3B1, and PK15 parent cells were 106.85, 103.63, and 104.74, respectively. The PK15-1A6 cell clone is more permissive to CSFV infection than the PK15 parent cells. The screened high-permissive cells will be useful for CSFV propagation and vaccine development in vitro, and facilitate research on the pathogenicity of CSFV.

High-throughput sequencing (HTS) identifies random viral fragments in environmental samples metagenomically. High reliability gains it broad application in virus evolution, host-virus interaction, and pathogenicity studies. Deep sequencing of field samples with content of host genetic material and bacteria often produces insufficient data for metagenomics and must be preceded by target enrichment. The main goal of the study was the evaluation of HTS for complete genome sequencing of field-case rabies viruses (RABVs). The material was 23 RABVs isolated mainly from red foxes and one European bat lyssavirus-1 isolate propagated in neuroblastoma cells. Three methods of RNA isolation were tested for the direct metagenomics and RABV-enriched approaches. Deep sequencing was performed with a MiSeq sequencer (Illumina) and reagent v3 kit. Bioinformatics data were evaluated by Kraken and Centrifuge software and de novo assembly was done with metaSPAdes. Testing RNA extraction procedures revealed the deep sequencing scope superiority of the combined TRIzol/column method. This HTS methodology made it possible to obtain complete genomes of all the RABV isolates collected in the field. Significantly greater rates of RABV genome coverages (over 5,900) were obtained with RABV enrichment. Direct metagenomic studies sequenced the full length of 6 out of 16 RABV isolates with a medium coverage between 1 and 71. Direct metagenomics gives the most realistic illustration of the field sample microbiome, but with low coverage. For deep characterisation of viruses, e.g. for spatial and temporal phylogeography during outbreaks, target enrichment is recommended as it covers sequences much more completely.

Objective: Present research aims to isolate, identify, and determine the virulence of the Streptococcus agalactiae (group B Streptococcus; GBS), isolated from popped eye disease affected Tilapia and Vietnamese Koi (V. Koi) fishes. Materials and Methods: A total of 330 fish samples were collected, of which Tilapia (n = 180) and V. Koi (n = 150), were collected from 35 affected ponds of four selected districts of Bangladesh. Isolation of the bacterium was done using different culture media (Nutrient broth, Plate count agar, Tryptic Soy Agar, and Blood agar), and identification by using various biochemical tests (con¬ventional and using API 20 Strep kit) and polymerase chain reaction (PCR) using primers against 16S rRNA gene of S. agalactiae. Antibiotic susceptibility of the bacteria was performed using seven different antibiotics disc (Tetracycline, Oxytetracycline, Chlortetracycline, Streptomycin, Ciprofloxacin, Gentamicin, and Neomycin). Virulence of the isolated S. agalactiae was determined by infecting healthy Tilapia and V. Koi fishes through experimental infection. Results: Isolated bacteria were found Gram-positive paired and chained cocci, β-hemolytic and non-motile. Findings of biochemical and serological tests indicate that the isolated bacterium belongs to Group B Streptococcus of Lancefield classification. PCR result also confirmed that the bacteria were S. agalactiae. The bacterial isolates possessed resistance property against all the seven antibiotics used in this study. The isolated GBS was found highly virulent and showed 80%90% mortality for Tilapia and V. Koi fishes in experimental infection within 16 days of post-infection. Conclusion: From the findings of this study, it may be concluded that isolated GBS from the Tilapia and V. Koi fishes were highly virulent and possessed multidrug-resistance properties. [J Adv Vet Anim Res 2021; 8(1.000): 14-23]

Objective: This study was carried out to assess the pathogenicity of local isolates of Mycoplasma ovipneumoniae and M. arginini in West African dwarf goats (kids) in Nigeria. Materials and methods: A total of 22 goats aged less than 1-year were purchased from markets. The goats were divided into six groups comprising of four experimental groups (EG; 4 in each) and two control groups (CG; 3 in each). The goats were fed ad libitum with standard diets and safe water. Groups EG1 and EG2 were infected with M. ovipneumoniae through trans-tracheal (TT) and intravenous (IV) routes, respectively, while those in groups EG3 and EG4 were infected with M. arginini through the same routes. Goats in groups CG1 and CG2 were inoculated with sterile Mycoplasma broth through TT and IV routes, respectively. In all cases, the amount of bacteria inoculated was 1.5x108 cells/mL. After the onset of the disease in goats, re-isolation of Mycoplasma was performed by culturing on mycoplasma agar supplemented with mycoplasma supplement G. The goats were monitored for 14 days post-infection (PI) to observe respiratory signs and mortality. Post-mortem (PM) examination was performed on each animal that died, while one surviving goat from each of the groups was sacrificed at 14 days PI for PM. After PM, histopathology was performed to observe the changes in tissues. Results: Cough and nasal discharges were observed in all the experimentally infected goats seven days PI. Mortalities were recorded in goats in EG1 (two goats), EG2 (one goats), EG3 (two goats) and EG4 (one goat). At PM, pneumonic lesions were observed in the lungs of all the experimentally infected goats. Conclusion: This study provides evidence that the local isolates of M. ovipneumoniae and M. arginini strains are pathogenic for goats in Nigeria. [J Adv Vet Anim Res 2016; 3(3.000): 242-251]

The current study was conducted to evaluate the pathogenicity and immunosuppressive effects of GM-97 strain of infectious bursal disease virus in commercial broiler chickens. A total of 500 broiler chickens were vaccinated with the virus through oral route at 10 and 17 days of age (102-103 EID50/dose). Chickens were also vaccinated with Newcastle disease virus (Hitchner B1) orally at 14 and 21 days old. Chickens were euthanized (at 12, 14, 16, 20, 23, 26 days of age) after measuring body weight. Bursa of Fabricius was examined for any gross lesion, weighed and processed for histological investigations. Bursa to body weight ratio and bursal lesion scoring were made to evaluate pathogenicity of the virus. Blood samples were analyzed for antibody response to ND vaccine virus using HI test. Results showed that the GM-97 strain of IBDV induced mild to moderate depletion of lymphoid cells in the center of bursal follicles and non-significant difference in bursa to body weight ratio amongst vaccinated and unvaccinated chickens. Chickens responded well to ND vaccine by mounting high level of serum NDV specific HI antibody titers. It can be concluded from the present study that GM-97 strain of IBDV has mild pathogenicity but is not immunosuppressive.

Objective: Salmonella is a widely-reported zoonotic bacterial pathogen and human infection is mostly attributed through direct or indirect contact with chickens. Salmonella Kentucky (S. Kentucky) is one of the motile serovars which has recently been identified from both poultry and human samples in Bangladesh. This study was conducted to assess its pathogenic potentials and shedding probability in backyard chicken. Materials and methods: We infected 22 backyard chickens orally, each with 106 cfu of Salmonella Kentuckey, which were then observed for 23 days to enlist clinical signs, gross and histo-pathological changes. Polymerase chain reaction (PCR) for Salmonella was applied on some representative samples to identify the presence of Salmonella. Results: Four chickens were sacrificed and the internal organs were examined to observe gross and microscopic tissue changes. Some reactive changes were seen in spleen during prolonged course of infection. The probability of S. Kentucky shedding was 77% (95%; CI 54-90%) on DPI 2, 41% (95%; CI 21-60%) on DPI 12 and 13% (95%; CI 3-31%) on DPI 21. The survival probability of the infected chickens was 50% (95%; CI 28-68%) on DPI 6, 32% (95%; CI 14-51%) on DPI 15 and 14% (95%; CI 3-31%) on DPI 23. Conclusion: Zoonotic S. Kentucky strain of human non-typhoidal clinical cases of gastroenteritis has potentials to produce clinical signs such as reduced feed uptake, watery or pasty fecal droppings and lesions, such as catarrhal enteritis and typhlitis. [J Adv Vet Anim Res 2018; 5(2.000): 196-203]

Objective: This study was conceived to investigate the pathogenicity of relapsed (Diminazene aceturate-resistant), field (original) and mixed (relapsed and field) isolates of Trypanosoma brucei brucei in rats.Materials and methods: Twenty eight healthy adult albino rats of both sexes weighing between 149-177 gm were used to compare the pathogenicity of relapsed, field and the mixed isolates of T. brucei brucei infections. The rats were separated into four groups (A-D); where, group A was kept as uninfected control, and group B was infected with 1x103 trypanosomes of the field isolate and 1x103 trypanosomes of the diminazene aceturate resistant isolate. The rats of groups C and D were infected with 1x106 trypanosomes of the diminazene aceturate-resistant isolate and 1x106 trypanosomes of the field isolate, respectively. Results: The infected rats became parasitemic within 4 to 8 days post-infection. The mean pre-patent periods (PP) were 4.1&plusmn;1.1, 6.0&plusmn;2.0 and 9.1&plusmn;1.1 days in groups B, C and D respectively, while the mean survival time (ST) in groups B, C and D were 21.4&plusmn;10.1, 27.1&plusmn;13.2 and 34.0 &plusmn;12.8 days, respectively. The PP and ST were shortest (P<0.05) in group B (mixed infections), and level of parasitemia was higher (P<0.05) in group B (mixed infections) as compared to groups C and D. The level of anemia was comparable (P>0.05) in groups C and D and more severe (P<0.05) in group B. Conclusion: Mixed infections exhibit shortest PP, ST, higher level of parasitemia and more severe anemia, and appear to be more pathogenic. [J Adv Vet Anim Res 2017; 4(1.000): 97-103]