Potency of bacterial sialidase Clostridium perfringens as antiviral of Newcastle disease infections using embryonated chicken egg in ovo model

Research (Published online: 06-08-2022)

1. Potency of bacterial sialidase Clostridium perfringens as antiviral of Newcastle disease infections using embryonated chicken egg in ovo model

Ryan Septa Kurnia, Simson Tarigan, Christian Marco Hadi Nugroho, Otto Sahat Martua Silaen, Lily Natalia, Fera Ibrahim, and Pratiwi Pudjilestari Sudarmono

Veterinary World, 15(8): 1896-1905

ABSTRACT

Background and Aim: Clostridium toxins are widely used as medicinal agents. Many active metabolic enzymes, including sialidase (neuraminidase), hyaluronidase, and collagenase, contribute to the mechanism of action of these toxins. Sialidase from Clostridium perfringens recognizes and degrades sialic acid receptors in the host cell glycoprotein, glycolipid, and polysaccharide complexes. Sialic acid promotes the adhesion of various pathogens, including viruses, under pathological conditions. This study aimed to investigate the potential of C. perfringens sialidase protein to inhibit Newcastle disease virus (NDV) infection in ovo model.

Materials and Methods: C. perfringens was characterized by molecular identification through polymerase chain reaction (PCR) and is cultured in a broth medium to produce sialidase. In addition, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis was conducted to characterize the sialidase protein. In contrast, enzymatic activity and protein concentration were carried out using a neuraminidase assay kit and Bradford to obtain suitable active substances. Furthermore, embryonated chicken egg models were used to observe the toxicity of several sialidase doses. Then, the hemagglutination (HA) titer was obtained, and absolute quantitative reverse transcription–PCR assay was performed to measure the viral replication inhibitory activity of sialidase against NDV.

Results: Each isolate had a specific sialidase gene and its product. The sialidase derived from C. perfringens could hydrolyze the sialic acid receptor Neu5Ac (2,6)-Gal higher than Neu5Ac (2,3)Gal in chicken erythrocytes, as observed by enzyme-linked lectin assay. A significant difference (p = 0.05) in the HA titer in the pre-challenge administration group at dosages of 375 mU, 187.5 mU, and 93.75 mU in the competitive inhibition experiment suggests that sialidase inhibits NDV reproduction. Quantification of infective viral copy confirmed the interference of viral replication in the pre-challenge administration group, with a significant difference (p = 0.05) at the treatment doses of 750 mU, 375 mU, and 46.87 mU.

Conclusion: The potency of sialidase obtained from C. perfringens was shown in this study, given its ability to reduce the viral titer and copy number in allantoic fluids without adversely impacting the toxicity of the chicken embryo at different concentrations.

Keywords: Clostridium perfringens, lectin, polysaccharide, sialidase, viral replication.

Occurrence, antimicrobial resistance, virulence, and biofilm formation capacity of Vibrio spp. and Aeromonas spp. isolated from raw seafood marketed in Bangkok, Thailand

Research (Published online: 31-07-2022)

37. Occurrence, antimicrobial resistance, virulence, and biofilm formation capacity of Vibrio spp. and Aeromonas spp. isolated from raw seafood marketed in Bangkok, Thailand

Sirijan Santajit, Thida Kong-ngoen, Witawat Tunyong, Pornpan Pumirat, Sumate Ampawong, Nitat Sookrung and Nitaya Indrawattana

Veterinary World, 15(7): 1887-1895

ABSTRACT

Background and Aim: Bacteria of the genera Vibrio and Aeromonas cause seafood-borne zoonoses, which may have a significant impact on food safety, economy, and public health worldwide. The presence of drug-resistant and biofilm-forming phenotypes in the food chain increases the risk for consumers. This study aimed to investigate the characteristics, virulence, biofilm production, and dissemination of antimicrobial-resistant pathogens isolated from seafood markets in Bangkok, Thailand.

Materials and Methods: A total of 120 retail seafood samples were collected from 10 local markets in Bangkok and peripheral areas. All samples were cultured and the Vibrio and Aeromonas genera were isolated using selective agar and biochemical tests based on standard protocols (ISO 21872-1: 2017). The antibiotic susceptibility test was conducted using the disk diffusion method. The presence of hemolysis and protease production was also investigated. Polymerase chain reaction (PCR) was used to determine the presence of the hlyA gene. Furthermore, biofilm formation was characterized by microtiter plate assay and scanning electron microscopy.

Results: The bacterial identification test revealed that 35/57 (61.4%) belonged to the Vibrio genus and 22/57 (38.6%) to the Aeromonas genus. The Kirby–Bauer test demonstrated that 61.4% of the isolates were resistant to at least one antibiotic and 45.61% had a high multiple antibiotic resistance index (≥0.2). PCR analysis indicated that 75.44% of the bacteria harbored the hlyA gene. Among them, 63.16% exhibited the hemolysis phenotype and 8.77% showed protease activity. The biofilm formation assay demonstrated that approximately 56.14% of all the isolates had the potential to produce biofilms. The moderate biofilm production was the predominant phenotype.

Conclusion: The results of this study provide evidence of the multiple drug resistance phenotype and biofilm formation capacity of Vibrio and Aeromonas species contaminating raw seafood. Effective control measures and active surveillance of foodborne zoonoses are crucial for food safety and to decrease the occurrence of diseases associated with seafood consumption.

Keywords: Aeromonas spp., biofilm formation, drug resistance, foodborne, seafood, vibriosis.

Feline upper respiratory tract disease – Computed tomography and laboratory diagnostic

Research (Published online: 30-07-2022)

36. Feline upper respiratory tract disease – Computed tomography and laboratory diagnostic

Armands Vekšins

Veterinary World, 15(7): 1880-1886

ABSTRACT

Background and Aim: Upper respiratory tract disease (URTD) is prevalent in cats, and diagnosis can be challenging. This study aimed to determine the most common causes of cat URTD in Latvia and describe computed tomography (CT) and laboratory diagnostic findings.

Materials and Methods: The present retrospective study included a total of 94 cats who were diagnosed with URTD. All cats underwent CT, and 50 of them had additional diagnostic tests, such as histology and respiratory infection polymerase chain reaction (PCR) testing.

Results: The most common CT finding was rhinosinusitis (55.32%) followed by nasal neoplasia (26.6%) and nasopharyngeal polyp (14.89%), but in three cats, a cause of respiratory symptoms was larynx neoplasia, nasal dermoid cyst, and an oronasal fistula. PCR test showed that the most cause of rhinosinusitis was Mycoplasma felis. Nasopharyngeal polyp as the primary diagnosis was identified in 14 cats from 3 months to 6 years, with an average age of 1.85 ± 1.915 years, and 54% of cats were female. Nasal neoplasia as a primary CT diagnosis was determined in 25 cats at the age of 5–18 years, with an average age of 10.56 ± 3.416 years. Histology diagnosis included four types of neoplasia – squamous cell carcinoma, sarcoma, adenocarcinoma, and aplastic carcinoma.

Conclusion: This study describes the most common CT and laboratory findings in cats with URTD. Included information will be helpful for general veterinary practitioners and researchers and will update their knowledge on feline URTD.

Keywords: computed tomography, feline, nasal neoplasia, Mycoplasma felis, rhinitis.

First study on the effect of transforming growth factor beta 1 and insulin-like growth factor 1 on the chondrogenesis of elephant articular chondrocytes in a scaffold-based 3D culture model

Research (Published online: 30-07-2022)

35. First study on the effect of transforming growth factor beta 1 and insulin-like growth factor 1 on the chondrogenesis of elephant articular chondrocytes in a scaffold-based 3D culture model

Siriwan Tangyuenyong, Patiwat Kongdang, Nutnicha Sirikaew and Siriwan Ongchai

Veterinary World, 15(7): 1869-1879

ABSTRACT

Background and Aim: Osteoarthritis (OA) is recognized as a degenerative joint disease that leads to chronic pain and low quality of life in animals. Captive elephants, the largest land mammals with a long lifespan, are more prone to develop OA due to restricted spaces and insufficient physical activity. This study aimed to investigate the effect of transforming growth factor-β1 (TGF-β1) and insulin-like growth factor 1 (IGF-1) on elephant chondrogenesis in a scaffold culture of articular chondrocytes.

Materials and Methods: Elephant chondrocytes-seeded gelatin scaffolds were cultured in chondrogenic media with or without 10 ng/mL of TGF-β1 or IGF-1 alone or 5–10 ng/mL of their combination for up to 21 days. The mRNA expression of cartilage-specific anabolic genes, ACAN and COL2A1, was analyzed using a real-time reverse transcription-polymerase chain reaction. The amounts of sulfated glycosaminoglycans (sGAGs) in conditioned media and contents in cultured scaffolds were determined through dimethylmethylene blue assay. Cell morphology, accumulation of proteoglycans, and details of the cultured scaffolds were determined using hematoxylin-eosin staining, safranin O staining, and scanning electron microscopy (SEM), respectively.

Results: TGF-β1 alone significantly upregulated ACAN gene expression but not COL2A1, while IGF-1 alone did not enhance both ACAN and COL2A1 genes. The combination significantly upregulated both mRNA expression levels of ACAN and COL2A1 gene at day 14. The sGAGs accumulation and contents in the treatment groups, except IGF-1 tended to be higher than the controls, concomitantly with the production of the extracellular matrix, showed the formation of a cartilage-like tissue through histological and SEM analyses.

Conclusion: Together, our results suggest that the single treatment of TGF-β1 has a selective effect on ACAN gene, while the combined growth factors seem to be an advantage on elephant chondrogenesis. This three-dimensional culture model is probably helpful for developing cartilage regeneration in vitro and is further applied in tissue engineering for OA treatment in vivo.

Keywords: articular chondrocyte, chondrogenesis, elephant, insulin-like growth factor 1, scaffold, transforming growth factor beta 1.

The first study on analysis of the codon usage bias and evolutionary analysis of the glycoprotein envelope E2 gene of seven Pestiviruses

Research (Published online: 29-07-2022)

34. The first study on analysis of the codon usage bias and evolutionary analysis of the glycoprotein envelope E2 gene of seven Pestiviruses

Mohammad Shueb, Shashanka K. Prasad, Kuralayanapalya Puttahonnappa Suresh, Uma Bharathi Indrabalan, Mallikarjun S. Beelagi, Chandan Shivamallu, Ekaterina Silina, Victor Stupin, Natalia Manturova, Shiva Prasad Kollur, Bibek Ranjan Shome, Raghu Ram Achar and Sharanagouda S. Patil

Veterinary World, 15(7): 1857-1868

ABSTRACT

Background and Aim: Pestivirus, a genus of the Flaviviridae family, comprises viruses that affect bovines, sheep, and pigs. Symptoms, including hemorrhagic syndromes, abortion, respiratory complications, and deadly mucosal diseases, are produced in infected animals, which cause huge economic losses to the farmers. Bovine viral diarrhea virus-1, bovine viral diarrhea virus-2, classical swine fever virus, border disease virus, Bungowannah, Hobi-like, and atypical porcine pestivirus belonging to the Pestivirus genus were selected for the study. This study aimed to estimate the codon usage bias and the rate of evolution using the glycoprotein E2 gene. Furthermore, codon usage bias analysis was performed using publicly available nucleotide sequences of the E2 gene of all seven Pestiviruses. These nucleotide sequences might elucidate the disease epidemiology and facilitate the development of designing better vaccines.

Materials and Methods: Coding sequences of the E2 gene of Pestiviruses A (n = 89), B (n = 60), C (n = 75), D (n = 10), F (n = 07), H (n = 52), and K (n = 85) were included in this study. They were analyzed using different methods to estimate the codon usage bias and evolution. In addition, the maximum likelihood and Bayesian methodologies were employed to analyze a molecular dataset of seven Pestiviruses using a complete E2 gene region.

Results: The combined analysis of codon usage bias and evolutionary rate analysis revealed that the Pestiviruses A, B, C, D, F, H, and K have a codon usage bias in which mutation and natural selection have played vital roles. Furthermore, while the effective number of codons values revealed a moderate bias, neutrality plots indicated the natural selection in A, B, F, and H Pestiviruses and mutational pressure in C, D, and K Pestiviruses. The correspondence analysis revealed that axis-1 significantly contributes to the synonymous codon usage pattern. In this study, the evolutionary rate of Pestiviruses B, H, and K was very high. The most recent common ancestors of all Pestivirus lineages are 1997, 1975, 1946, 1990, 2004, 1990, and 1990 for Pestiviruses A, B, C, D, F, H, and K, respectively. This study confirms that both mutational pressure and natural selection have played a significant role in codon usage bias and evolutionary studies.

Conclusion: This study provides insight into the codon usage bias and evolutionary lineages of pestiviruses. It is arguably the first report of such kind. The information provided by the study can be further used to elucidate the respective host adaptation strategies of the viruses. In turn, this information helps study the epidemiology and control methods of pestiviruses.

Keywords: codon usage bias, evolutionary analysis, Flaviviridae, glycoprotein E2, India, Pestivirus.

Difference in cytological findings of healthy and conjunctivitis/keratoconjunctivitis affected canine eyes between variably experienced evaluators

Research (Published online: 28-07-2022)

33. Difference in cytological findings of healthy and conjunctivitis/keratoconjunctivitis affected canine eyes between variably experienced evaluators

Inese Berzina, Anastasija Terentjeva and Liga Kovalcuka

Veterinary World, 15(7): 1852-1856

ABSTRACT

Background and Aim: Cytology investigations are a frequent part of ophthalmological examination. We aimed to assess whether the cytological findings of healthy and conjunctivitis/keratoconjunctivitis samples differed based on the evaluator's experience.

Materials and Methods: A study evaluated healthy eyes (n = 40) and eyes affected with keratoconjunctivitis and/ or conjunctivitis (n = 28) in dogs. An ophthalmological examination was performed before sampling the eyes using a sterile cotton swab. An evaluator with theoretical experience and one with undergone clinical pathology residency training performed cytology blinded to the clinical findings.

Results: In the healthy eyes group, the agreement between the evaluators for cellularity was nonexistent, while that for cell preservation and mucus content was fair. In the affected eyes group, the agreement for cellularity and mucus content was moderate, while that for cell preservation was fair. The inadequate sample rate differed significantly between the two evaluators in the healthy eyes group (p = 0.006) but not in the affected eyes group (p = 0.083). Bacterial presence was detected by both evaluators, and the findings did not differ statistically from the bacteriology results (p = 0.05). Significant variations were noted in the differential cell count; the mean count of the superficial epithelial cells and goblet cells of the healthy eyes group (p < 0.05) and that of the basal/intermediate cells and neutrophils of the affected eyes (p < 0.05) showed significant differences.

Conclusion: The evaluator's experience significantly affected the differential cell count in both the healthy and affected eyes groups. Neutrophil degeneration was not observed by the less experienced evaluator, whereas bacteria were detected equally well by both the evaluators.

Keywords: conjunctival cytology, microscopy experience, neutrophil morphology.

Percent positivity and phylogenetic analysis of Mycoplasma gallisepticum and Mycoplasma synoviae in commercial poultry from the different States of India

Research (Published online: 28-07-2022)

32. Percent positivity and phylogenetic analysis of Mycoplasma gallisepticum and Mycoplasma synoviae in commercial poultry from the different States of India

Pranoti Giram, Pankhudi Bhutada, Chhagan Prajapati, Santosh S. Koratkar, Sachin Patil, Devender Hooda, Vinay Rale, and Satish S. Tongaonkar

Veterinary World, 15(7): 1843-1851

ABSTRACT

Background and Aim: The Indian and global poultry industries suffer significant economic losses due to Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) infections, which adversely affect egg production, hatchability, weight gain, and feed efficiency in farms, thus decreasing the overall production efficiency. This study aimed to determine the percent positivity and phylogenetic analysis of MG, MS, and co-infection of both mycoplasmas in commercial poultry farms across different states of India from 2017 to 2021.

Materials and Methods: A total of 3620 tracheal or choacal swabs were collected from breeder and layer farms showing clinical signs of avian mycoplasma infections from commercial poultry farms across India, and the percent positivity for MG, MS, and co-infection of both mycoplasmas were determined by Polymerase chain reaction using the 16S rRNA and vlhA genes amplification, respectively. Phylogenetic analysis was carried out by sequencing the mgc2 and vlhA genes of 2 samples of MG and 24 samples of M. synoviae to gain insight into the genetic variability of Indian strains. The data were then compared with other Indian strains, vaccines strains, and strains from other countries.

Results: Our data shows the percent positivity of MG, MS, and co-infection of both MG and MS was 6.43%, 23.61%, and 15.49%, respectively. The phylogenetic relationship between MG and MS was determined using the vlhA and mgc2 genes, revealing two samples of MG and 24 samples of MS clustered with other Indian strains. M. synoviae MSM22 and previously studied M. synoviae MGS 482 clustered with vaccine strain M. synoviae MS-H.

Conclusion: Mycoplasma synoviae infections in breeder, layer, and in both is predominant compared to MG across the states investigated in India. Sequenced samples of MG and MS showed evolutionary relationships with the previously studied Indian strains of MG and MS. These findings support our view that monitoring chickens for avian mycoplasma infections are of paramount significance. It further lends credence to the contention that such information will pave the way for the development of a home-grown vaccination control program and thus safeguard the poultry sector against mycoplasma infections.

Keywords: epidemiology, India, Mycoplasma gallisepticum, Mycoplasma synoviae, phylogeny, polymerase chain reaction.