Toxicity test of flavonoid compounds from the leaves of Dendrophthoe pentandra (L.) Miq. using in vitro culture cell models

Research (Published online: 21-12-2022)

18. Toxicity test of flavonoid compounds from the leaves of Dendrophthoe pentandra (L.) Miq. using in vitro culture cell models

Mochamad Lazuardi, Suharjono Suharjono, Chi-Hsien Chien, Jie-Long He, Chi-Wen Lee, Chia-Kang Peng, Mohammad Sukmanadi, Rahmi Sugihartuti, and Lilik Maslachah

Veterinary World, 15(12): 2896-2902

ABSTRACT

Background and Aim: The flavonoids from mistletoe are thought to have antimicrobial action. This encouraging finding supports the benefits of medicinal plants as a substitute for synthetic antimicrobials, thus promoting healthy lifestyles. In contrast, it is known that the use of topical drug formulations made from flavonoids of mistletoe (Dendrophthoe pentandra (L.) Miq. Loranthaceae) with Indonesian name, Benalu duku (BD) is required in skin cell irritation. This study aimed to assess the toxic effects of the flavonoid substances of BD, as an initial screening.

Materials and Methods: A myeloma cell line was cultured in Roswell Park Memorial Institute medium, and the Baby Hamster Kidney clone 12 (BHK21) cell line was cultured in Dulbecco's Modified Eagle's Medium from stock (±9 × 107 cells/mL), and 1.2 mL of culture were distributed into each well of a microtiter plate. Subsequently, 0.2 mL of serially diluted flavonoid compounds (0.5–3 μg/mL) were added to 12 wells for each concentration, as trial groups (including control groups), followed by a 2-day incubation. Observations were performed based on the cytopathic effect (CPE) using an inverted microscope at a magnification of 100×.

Results: Cytopathic effect was detected on the microtiter plate wells for the groups of myeloma and BHK21 cells at a flavonoid concentration of 0.5 μg/mL–3 μg/mL.

Conclusion: Flavonoid compounds from BD were safely used for topical treatment of cancer cells at a concentration <2.491 μg/mL, whereas for non-cancerous cells, a concentration <2.582 μg/mL was sufficient (p < 0.05).

Keywords: antimicrobial, Baby Hamster Kidney clone 12 cell culture, cytopathic effect, healthy lifestyle, myeloma cell culture.

Effect of follicle-stimulating hormone and luteinizing hormone levels on egg-laying frequency in hens

Research (Published online: 20-12-2022)

17. Effect of follicle-stimulating hormone and luteinizing hormone levels on egg-laying frequency in hens

Ragil Angga Prastiya, Sri Pantja Madyawati, Sera Yunita Sari, and Aras Prasetiyo Nugroho

Veterinary World, 15(12): 2890-2895

ABSTRACT

Background and Aim: Gonadotropins, for example, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), are hormones that affect the reproductive process. In hens, optimal levels of FSH and LH can stimulate follicle growth fairly rapidly and thereby increase egg production through follicle development and increased ovulation. Follicle-stimulating hormone acts in the early stages of follicular growth, whereas LH acts on pre-ovulatory follicles. Normal follicular growth is the result of the complementary action of FSH and LH. Low FSH and LH levels result in the formation of follicles but a lack of egg production in chickens. This study aimed to investigate FSH and LH hormone levels from layer chickens with different egg-laying frequencies.

Materials and Methods: Fifty blood serum samples were collected from 54-week-old ISA brown strain hens that were divided into five groups (with 10 hens per group) as follows: Hens that lay eggs (i) every day, (ii) once every 2 days, (iii) twice every 3 days, (iv) 3 times every 4 days, and (v) hens that do not lay eggs. Follicle-stimulating hormone and LH levels were measured in samples using an enzyme-linked immunosorbent assay, and the data were analyzed using multivariate analysis of variance.

Results: Follicle-stimulating hormone levels were significantly associated with the frequency of egg laying in ISA brown strain hens (p < 0.05); the highest FSH level (869.005 ± 149.194 pg/mL) was found in hens that lay eggs every day. In contrast, the highest LH level (51.386 ± 2.410 mIU/mL) was found in hens that lay eggs every 2 days.

Conclusion: High level of FSH (869.005 ± 149.194 pg/mL) was associated with a high frequency of egg laying (every day) in ISA brown strain hens, and LH level of around 30.406 pg/mL was associated with daily egg laying in these hens.

Keywords: egg-laying frequency, enzyme-linked immunosorbent assay, gonadotropin hormone, laying hens, reproductive health.

Molecular detection and phylogeny of Ehrlichia canis and Anaplasma platys in naturally infected dogs in Central and Northeast Thailand

Research (Published online: 20-12-2022)

16. Molecular detection and phylogeny of Ehrlichia canis and Anaplasma platys in naturally infected dogs in Central and Northeast Thailand

Andaman Purisarn, Sakulchit Wichianchot, Cherdsak Maneeruttanarungroj, Bandid Mangkit, Wuttinun Raksajit, Sarawan Kaewmongkol, Thitichai Jarudecha, Wanat Sricharern, and Rucksak Rucksaken

Veterinary World, 15(12): 2877-2889

ABSTRACT

Background and Aim: Ehrlichia canis and Anaplasma platys are tick-borne, Gram-negative bacteria that cause canine monocytic ehrlichiosis and canine cyclic thrombocytopenia, respectively. These diseases are of great importance and are distributed globally. This study aimed to create new primers for the identification of E. canis and A. platys in naturally infected dogs using polymerase chain reaction (PCR), DNA sequencing, and phylogenetic analysis using the 16S rDNA and gltA genes.

Materials and Methods: In total, 120 blood samples were collected from dogs in three different locations (Saraburi, Buriram, and Nakhon Ratchasima provinces) in Central and Northeast Thailand. The molecular prevalence of E. canis and A. platys was assessed using PCR targeting the 16S rDNA and gltA genes. All positive PCR amplicons were sequenced, and phylogenetic trees were constructed based on the maximum likelihood method.

Results: Ehrlichia canis had an overall molecular prevalence of 15.8% based on the 16S rDNA gene, compared to 8.3% based on the gltA gene. In addition, the overall molecular prevalence of A. platys using the 16S rDNA gene was 10.8%, while the prevalence rate was 5.8% using the gltA gene. Coinfection was 0.8% in Saraburi province. The partial sequences of the 16S rDNA and gltA genes of E. canis and A. platys in dogs in Central and Northeast Thailand showed 96.75%–100% identity to reference sequences in GenBank. Phylogenetic analysis of the 16S rDNA and gltA genes revealed that E. canis and A. platys sequences were clearly grouped into their own clades.

Conclusion: This study demonstrated the molecular prevalence of E. canis and A. platys in Central and Northeast Thailand. The 16S rDNA and gltA genes were useful for the diagnosis of E. canis and A. platys. Based on the phylogenetic analysis, the partial sequences of the 16S rDNA and gltA genes in E. canis and A. platys were related to prior Thai strains and those from other countries.

Keywords: 16S rDNA gene, Anaplasma platys, Ehrlichia canis, gltA gene, phylogenetic analysis.

Isolation of multidrug-resistant Escherichia coli and Salmonella spp. from sulfonamide-treated diarrheic calves

Research (Published online: 16-12-2022)

15. Isolation of multidrug-resistant Escherichia coli and Salmonella spp. from sulfonamide-treated diarrheic calves

Mohammad Amdadul Haque, Muhammad Tofazzal Hossain, Md. Shafiqul Islam, Md. Zahorul Islam, Purba Islam, Sourendra Nath Shaha, Mahmudul Hasan Sikder, and Kazi Rafiq

Veterinary World, 15(12): 2870-2876

ABSTRACT

Background and Aim: The bovine industry is threatened by one of the most serious and deadly enteric diseases, calf diarrhea, particularly in developing nations like Bangladesh. In this context, bacterial resistance to antimicrobial drugs and its detrimental consequences have become a critical public health issue that is difficult to address globally. This study aimed to isolate and identify Escherichia coli and Salmonella spp. with their antibiogram and antibiotic resistance gene detection from sulfonamide-treated diarrheic calves.

Materials and Methods: Twelve diarrheic calves suffering from calf diarrhea in a dairy farm were selected and a total of 36 fecal samples were aseptically collected directly from rectum before, during, and at the end of treatment for each calf to determine the total viable count, total E. coli count and total Salmonella count. A polymerase chain reaction was used for the specific detection of E. coli and Salmonella genus targeting fliC and invA genes, respectively. Antibiotic sensitivity test of the isolated E. coli and Salmonella spp. were performed by the disk diffusion method for eight commonly used antibiotics.

Results: A total of 36 E. coli (100%) and 12 Salmonella spp. (33%) were isolated from the samples and were confirmed by polymerase chain reaction. Total viable count was found to be ranged from 35 × 107 to 99 × 1010 colony-forming unit (CFU)/g fecal sample before starting sulfonamide treatment, 34 × 105 to 25 × 1010 CFU/g during treatment with sulfonamide, and 48 × 103 to 69 × 1010 CFU/g immediately after completion of sulfonamide treatment. Total E. coli count was found to be ranged from 4 × 104 to 36 × 1010 CFU/g, 24 × 104 to 23 × 108 CFU/g, and 13 × 104 to 85 × 1010 CFU/g, whereas total Salmonella count was found to be ranged from 16 × 106 to 18.5 × 1011 CFU/g, 15 × 104 to 44 × 107 CFU/g, and 13.2 × 105 to 21 × 1010 CFU/g fecal sample before starting sulfonamide treatment, during treatment with sulfonamide immediately after completion of sulfonamide treatment, respectively. The in vitro antibiotic sensitivity test showed that all the E. coli and Salmonella spp. isolated from diarrheic calves (100%) contained multidrug-resistant (MDR) phenotypes. Escherichia coli isolates were found 100% resistant to amoxicillin (AMX), cefuroxime, cephalexin (CN), erythromycin (ERY), and tetracycline (TET); whereas 94.4%, 86.1%, and 77.8% isolates were resistant to doxycycline (DOX), moxifloxacin (MOF), and gentamycin (GEN), respectively. In case of Salmonella isolates, all were found 100% resistant to AMX, CN, and ERY; whereas 91.7% of resistance was observed for DOX, MOF, cefuroxime, GEN, and TET. Based on the molecular screening of the antibiotic resistance genes, tetA gene was present in 83.3% of the isolated E. coli and 75% of the isolated Salmonella strains, whereas 83.3% E. coli and 79.2% Salmonella isolates contained blaTEM gene.

Conclusion: These findings suggest that MDR E. coli and Salmonella spp. might be responsible for calf scouring, which is challenging to treat with antibiotics or sulfonamide drugs alone. Therefore, it is important to check the antibiotic sensitivity pattern to select a suitable antibiotic for the treatment of calf scoring. A suitable antibiotic or combination of an antibiotic and sulfonamide could be effective against E. coli and Salmonella spp. responsible for calf scouring.

Keywords: antimicrobial resistance, calf, diarrhea, Escherichia coli, multidrug-resistant, Salmonella spp.

Selection and characterization of bacteriophages specific to Salmonella Choleraesuis in swine

Research (Published online: 16-12-2022)

14. Selection and characterization of bacteriophages specific to Salmonella Choleraesuis in swine

Pattaraporn Sriprasong, Napakhwan Imklin, and Rujikan Nasanit

Veterinary World, 15(12): 2856-2869

ABSTRACT

Background and Aim: Salmonella Choleraesuis is the most common serotype that causes salmonellosis in swine. Recently, the use of bacteriophages as a potential biocontrol strategy has increased. Therefore, this study aimed to isolate and characterize bacteriophages specific to S. Choleraesuis associated with swine infection and to evaluate the efficacy of individual phages and a phage cocktail against S. Choleraesuis strains in simulated intestinal fluid (SIF).

Materials and Methods: Three strains of S. Choleraesuis isolated from pig intestines served as host strains for phage isolation. The other 10 Salmonella serovars were also used for the phage host range test. The antibiotic susceptibility of the bacterial strains was investigated. Water samples from natural sources and drain liquid from slaughterhouses were collected for phage isolation. The isolated phages were characterized by determining the efficiency of plating against all Salmonella strains and the stability at a temperature range (4°C–65°C) and at low pH (2.5–4.0) in simulated gastric fluids (SGFs). Furthermore, morphology and genomic restriction analyses were performed for phage classification phages. Finally, S. Choleraesuis reduction in the SIF by the selected individual phages and a phage cocktail was investigated.

Results: The antibiotic susceptibility results revealed that most Salmonella strains were sensitive to all tested drugs. Salmonella Choleraesuis KPS615 was multidrug-resistant, showing resistance to three antibiotics. Nine phages were isolated. Most of them could infect four Salmonella strains. Phages vB_SCh-RP5i3B and vB_SCh-RP61i4 showed high efficiency in infecting S. Choleraesuis and Salmonella Rissen. The phages were stable for 1 h at 4°C–45°C. However, their viability decreased when the temperature increased to 65°C. In addition, most phages remained viable at a low pH (pH 2.5–4.0) for 2 h in SGF. The efficiency of phage treatment against S. Choleraesuis in SIF showed that individual phages and a phage cocktail with three phages effectively reduced S. Choleraesuis in SIF. However, the phage cocktails were more effective than the individual phages.

Conclusion: These results suggest that the newly isolated phages could be promising biocontrol agents against S. Choleraesuis infection in pigs and could be orally administered. However, further in vivo studies should be conducted.

Keywords: antimicrobials, bacteriophage, biocontrol, Salmonella Choleraesuis, swine.

Estimation of the herd-level basic reproduction number for African swine fever in Vietnam, 2019

Research (Published online: 15-12-2022)

13. Estimation of the herd-level basic reproduction number for African swine fever in Vietnam, 2019

Thi Ngan Mai, Thanh Trung Nguyen, Viet Anh Vu, Thi Ngoc Vu and Thi My Le Huynh

Veterinary World, 15(12): 2850-2855

ABSTRACT

Background and Aim: African swine fever (ASF) is a notifiable viral disease of pigs and wild boars that causes severe economic losses to the swine industry. The pig industry in Vietnam was recently attacked by the ASF virus (ASFV) for the first time in history. However, we lack information regarding the transmissibility of ASF within indoor production systems communities, such as those in Vietnam. Therefore, we aimed to estimate the basic reproduction number (R0) for ASF during the early stages of transmission between farms in indoor production system communities from local and national data in Vietnam.

Materials and Methods: The linear regression model approach for the susceptible infectious method was used in this study to estimate the transmission rate and, consequently, the R0 value.

Results: The R0 values between-farm of ASF ranged from 1.41 to 10.8 in different scenarios of infectious period duration, from 15 to 30 days at the national and local levels.

Conclusion: These results help to understand the scale and speed of ASF infection in Vietnam and to provide a scientific basis to implement control measures to restrict the spread of ASFV in other locations.

Keywords: African swine fever, basic reproduction number, indoor production, Vietnam.

Prevalence of Toxoplasma gondii, Leptospira spp., and Coxiella burnetii-associated antibodies in dairy cattle with reproductive disorders

Research (Published online: 15-12-2022)

12. Prevalence of Toxoplasma gondii, Leptospira spp., and Coxiella burnetii-associated antibodies in dairy cattle with reproductive disorders

V. Balamurugan, K. Vinod Kumar, Anusha Alamuri, P. P. Sengupta, G. Govindaraj, and B. R. Shome

Veterinary World, 15(12): 2844-2849

ABSTRACT

Background and Aim: In cattle dairy farms, abortions and other reproductive problems due to major infectious diseases are overlooked, and identifying their causative agents is very challenging without a confirmatory diagnosis. Further, a prevalence study in animals will provide important hints of pathogen reservoirs and provide necessary direction to disease burden with appropriate control and biosecurity measures at the farm level. This study aimed to estimate the prevalence of Toxoplasma gondii antibodies in dairy cattle associated with reproductive problems along with coexisting antibodies against abortifacient zoonotic (Coxiella burnetii and Leptospira spp.) pathogens.

Materials and Methods: Cattle sera (n = 246) from dairy farms (n = 35) situated in different locations in India were screened for anti-T. gondii and C. burnetii antibodies with enzyme-linked immunoassay and Leptospira spp. antibodies with microscopic agglutination test.

Results: The overall prevalence of 11.4% (95% confidence intervals [CIs]: 7.99%–15.96%) antibodies in cattle associated with reproductive problems (p < 0.021) with farm-level seropositivity of 43% was observed. Further, on analysis of screened sera, 49.8% (95% CI: 42.6%–55%) and 77.6% (95% CI: 72%–82.4%) of samples were found to be positive for C. burnetii and Leptospira spp. antibodies, respectively. Moreover, the seropositivity of 91.9% (226/246) for at least one of the screened zoonotic pathogens was observed, indicating antibodies against either of these organisms in association with reproductive disorders (p < 0.005). The percentage of cattle found to have T. gondii antibodies was only 1.8%, whereas 11.5% and 41.6% of cattle were found to have C. burnetii and Leptospira spp. antibodies, respectively. Nevertheless, the predominantly mixed infections observed were of Leptospira and C. burnetii (34.5%), followed by all three infections (4.9%); toxoplasmosis and leptospirosis (3.5%); and toxoplasmosis and Q fever (2.2%).

Conclusion: The serological detection of antibodies against these pathogens in cattle may have significant implications for the livestock industry and public health, suggesting the need for continuous surveillance and monitoring of these infections to prevent their spread.

Keywords: dairy cattle, leptospirosis, Q fever, seroprevalence, toxoplasmosis.

The reversible effects of gossypol toxicity on male pigeons' reproductive performance

Research (Published online: 13-12-2022)

11. The reversible effects of gossypol toxicity on male pigeons' reproductive performance

Suwarak Wannaratana, Wijit Banlunara, Kaj Chokeshaiusaha, and Thanida Sananmuang

Veterinary World, 15(12): 2836-2843

ABSTRACT

Background and Aim: Gossypol, a cotton seed derivative, is well known for its reversible antifertility in male reproduction across species. Its antifertility and reversibility effects on male reproductive function vary among species in dose-and time-dependent manners. In this study, the antifertility potential of gossypol in pigeons was evaluated for the first time to determine whether it might be used as a dietary supplement for pigeon population control.

Materials and Methods: Male pigeons were assigned into three experimental groups: The gossypol-treated group (n = 12), the sham control group (n = 6), and the negative control group (n = 6). There were two experimental periods: A gossypol-feeding period of 28 days and a gossypol-free period of 28 days. During the gossypol-feeding period, birds in the gossypol-treated group were fed 4 mg of gossypol extract per day. Birds in the sham control group were fed 0.5 mL of mixed ethanol and sunflower oil, while those in the negative control group were fed 0.5 mL of phosphate buffer saline. After the gossypol-feeding phase was completed, all remaining pigeons in all groups continued to receive their regular diet for an additional 28 days (gossypol-free phase). The body weight and semen quality of the birds in the experimental groups were compared to evaluate gossypol's antifertility effect.

Results: In the gossypol-treated group as compared to the control groups, the percentages of sperm motility and viability were significantly lower at 21 days, and the percentage of normal sperm morphology was significantly lower at 28 days during the gossypol-feeding period. After gossypol withdrawal, these antifertility effects were resumed and reached a comparable semen quality to the control groups within 14 days.

Conclusion: Gossypol supplementation (4 mg/day for 28 days) could lower male pigeons' reproductive performance in terms of sperm motility, viability, and sperm morphology. Such infertility was, however, reversible within 14 days after gossypol withdrawal without any side effects on the pigeons, suggesting its application as a safe contraceptive feeding for male pigeons.

Keywords: gossypol, pigeon, sperm morphology, sperm motility, sperm viability.

Seroprevalence of nasal myiasis in camels determined by indirect enzyme-linked immunosorbent assay utilizing the most diagnostic Cephalopina titillator larval antigens

Research (Published online: 13-12-2022)

10. Seroprevalence of nasal myiasis in camels determined by indirect enzyme-linked immunosorbent assay utilizing the most diagnostic Cephalopina titillator larval antigens

Noha M. F. Hassan, Doaa Sedky, Nadia M. T. Abu El Ezz, and Eman E. El Shanawany

Veterinary World, 15(12): 2830-2835

ABSTRACT

Background and Aim: Nasal myiasis is a serious parasitic disease among camels caused by Cephalopina titillator larvae that negatively affect animal health and production globally. The diagnosis of the infestation relies on postmortem examination of the head region, which considers a cause impeding treatment of live animals and may be misdiagnosed as central nervous system disorders. This study aimed to identify the most diagnostic larval antigen with the capacity for monitoring C. titillator infestation, and to estimate the seroprevalence of nasal myiasis in camels in Egypt, using indirect enzyme-linked immunosorbent assay (ELISA).

Materials and Methods: Three hundred and six male camels of Egyptian and Sudanese breeds, aged 2–5 years, were clinically evaluated for respiratory and/or nervous disorders in Cairo Governorate, Egypt. At the time of slaughter, blood samples were collected from all examined animals. The postmortem examination of 38 animals was conducted. Salivary glands, hemolymph, and somatic antigens were extracted from the second and third larval instars.

Results: The results revealed that the salivary gland antigen was the most potent antigen in detecting C. titillator specific total IgG antibodies compared to haemolymph and crude somatic antigens. Using receiver-operating characteristic curves and area under the curve, the salivary gland antigen had a sensitivity of 91.67% and a specificity of 92.31%, respectively. It has the highest positive predictive value, 95.7%, and negative predictive value, 85.7%. However, using somatic and hemolymph antigens revealed a sensitivity of 79.17% and 70.83% and a specificity of 76.9% and 84.6%, respectively. There was complete concordance between ELISA results and autopsy findings (true positive). One hundred and forty out of 306 (45.8%) camel serum samples were found to contain C. titillator.

Conclusion: This study demonstrated that salivary gland antigen is more effective than somatic and hemolymph antigens in accurately detecting nasal myiasis in camels. In addition, determining the seroprevalence of nasal myiasis with the salivary gland antigen through indirect ELISA revealed that it is a prevalent disease among camels in Egypt. Periodic surveillance of the C. titillator prevalence is necessary for effective management and control measures.

Review of pork and pork products as a source for transmission of methicillin-resistant Staphylococcus aureus

Review (Published online: 12-12-2022)

8. Review of pork and pork products as a source for transmission of methicillin-resistant Staphylococcus aureus

Aswin Rafif Khairullah, Sri Agus Sudjarwo, Mustofa Helmi Effendi, Sancaka Cashyer Ramandinianto, Maria Aega Gelolodo, Agus Widodo, Katty Hendriana Priscilia Riwu, and Dyah Ayu Kurniawati

International Journal of One Health, 8(2): 167-177

ABSTRACT

Methicillin-resistant Staphylococcus aureus (MRSA) is an opportunistic bacterium that can cause infection in animals and humans. Recently, MRSA from food-producing or farm animals has been identified as livestock-associated MRSA (LA-MRSA). The spread of LA-MRSA is particularly found in pork and pork products because LA-MRSA has been widely known to infect pigs. The most common type of LA-MRSA identified in pork and pork products is the clonal complex LA-MRSA 398 (LA-MRSA CC398). The MRSA strains on the surface of pork carcasses can be spread during the handling and processing of pork and pork products through human hands, cutting tools, and any surface that comes into direct contact with pork. Food infection is the main risk of MRSA in pork and pork products consumed by humans. Antibiotics to treat food infection cases due to MRSA infection include vancomycin and tigecycline. The spread of MRSA in pork and pork products is preventable by appropriately cooking and cooling the pork and pork products at temperatures above 60°C and below 5°C, respectively. It is also necessary to take other preventive measures, such as having a clean meat processing area and disinfecting the equipment used for processing pork and pork products. This review aimed to explain epidemiology, transmission, risk factors, diagnosis, public health consequences, treatment of food poisoning, and preventing the spread of MRSA in pork and pork products.

Keywords: antibiotics, livestock, methicillin-resistant Staphylococcus, pork products.

Multidrug resistance-encoding gene in Citrobacter freundii isolated from healthy laying chicken in Blitar District, Indonesia

Research (Published online: 12-12-2022)

7. Multidrug resistance-encoding gene in Citrobacter freundii isolated from healthy laying chicken in Blitar District, Indonesia

Adiana Mutamsari Witaningrum, Freshinta Jellia Wibisono, Dian Ayu Permatasari, Mustofa Helmi Effendi, and Emmanuel Nnabuike Ugbo

International Journal of One Health, 8(2): 161-166

ABSTRACT

Background and Aim: The increasing prevalence of resistance (MDR) of Enterobacteriaceae in Indonesia has caused concern regarding human health. Citrobacter freundii reportedly targets the gastrointestinal tract of animals and is a common cause of foodborne diseases associated with diarrhea, peritonitis, meningitis, brain abscess, bacteremia, and urinary tract infection. This study aimed to estimate the prevalence of MDR and the presence of Class 1 integron-encoding genes in C. freundii isolates obtained from cloacal swabs of healthy laying chickens in Blitar district, Indonesia.

Materials and Methods: One hundred and sixty-five cloacal swab samples were collected from 33 farms in Blitar over a period of 4 months. Standard microbiological techniques such as bacterial culture in MacConkey agar, Simmons citrate agar, and triple sugar iron agar and biochemical tests such as the indole test were performed to identify the isolates. The antibiotic sensitivity patterns of C. freundii isolates were determined by the disk diffusion method, and MDR-encoding genes (Class 1 integron) were detected by polymerase chain reaction (PCR).

Results: Out of 165 cloacal swab samples, 7 (4.24%) were positive for C. freundii. Citrobacter freundii was highly resistant to erythromycin (71.43%) and moderately to streptomycin, tetracycline, and trimethoprim-sulfamethoxazole (all 42.86%); however, it showed low resistance to ampicillin (28.57%). All isolates were found to exhibit MDR. Only 1 (14.29%) of the seven C. freundii isolates harbored a Class 1 integron gene. This study revealed that Class 1 integron-encoding genes have a low prevalence in C. freundii isolated from healthy laying chickens in Blitar, Indonesia.

Conclusion: Poultry animals can play a role in the transmission of resistance genes to humans due to the MDR of Enterobacteriaceae, including C. freundii in the intestines.

Keywords: Citrobacter freundii, Class 1 integron gene, human health, laying chickens, multidrug resistance.

Bacteriophage efficacy in controlling swine enteric colibacillosis pathogens: An in vitro study

Research (Published online: 12-12-2022)

9. Bacteriophage efficacy in controlling swine enteric colibacillosis pathogens: An in vitro study

Techaphon Songphasuk, Napakhwan Imklin, Pattaraporn Sriprasong, Yonlayong Woonwong, Rujikan Nasanit, and Somchai Sajapitak

Veterinary World, 15(12): 2822-2829

ABSTRACT

Background and Aim: Swine enteric colibacillosis caused by Escherichia coli is a major problem in the swine industry, causing diarrhea among swine and resulting in substantial financial losses. However, efforts to counter this disease are impeded by the increase in antimicrobial resistance (AMR) worldwide, so intensive research is being conducted to identify alternative treatments. This study isolated, characterized, and evaluated the efficacy of bacteriophages to control pathogens causative of swine enteric colibacillosis.

Materials and Methods: Five sewage samples were collected from different areas of a swine farm in Suphanburi province, Thailand and the bacteriophages were enriched and isolated, followed by purification by the agar overlay method using E. coli RENR as the host strain. The selected phages were characterized by evaluating their morphology, while their specificity was verified by the host range test. The efficiency of plating and multiplicity of infection (MOI) were also determined.

Results: Four selected phages, namely, vB_Eco-RPNE4i3, vB_Eco-RPNE6i4, vB_Eco-RPNE7i1, and vB_Eco-RPNE8i3, demonstrated different patterns of host range and phage efficiency. They significantly decreased E. coli concentration at the tested MOIs (0.01–100) from 1 h onward. However, bacterial regrowth was observed in all phage treatments.

Conclusion: This study shows the potential of using phages as an alternative treatment for swine enteric colibacillosis. The obtained results demonstrated that the selected phages had a therapeutic effect against pathogens causative of swine enteric colibacillosis. Therefore, phages could be applied as an alternative treatment to control specific bacterial strains and reduce AMR arising from the overuse of antibiotics.

Keywords: bacteriophages, colibacillosis, diarrhea, Escherichia coli, swine.

Molecular characterization of Newcastle disease virus vaccines in Nigeria

Research (Published online: 10-12-2022)

8. Molecular characterization of Newcastle disease virus vaccines in Nigeria

Mohammed Usman Sajo, Lawal Sa'idu, Maman Moutari Souley, and Olusegun Adesina Fagbohun

Veterinary World, 15(12): 2816-2821

ABSTRACT

Background and Aim: Newcastle disease (ND) caused by ND virus (NDV) is a serious impediment to effective poultry production in developing countries such as Nigeria. Despite employing vaccination and other control measures to curtail this disease, its severe forms still persist. This study aimed to confirm the virus strains in the NDV vaccine brands commonly used in Nigeria.

Materials and Methods: We employed reverse-transcription polymerase chain reaction (RT-PCR), sequencing, and sequence analysis to characterize NDV strains in four NDV vaccines commonly used in Nigeria. Fragments of 300 bp from NDV fusion genes from the vaccines were amplified. Polymerase chain reaction products were sequenced and analyzed using multiple sequence alignment and phylogenetic analyses to characterize the vaccine viruses as pathotypes.

Results: All the vaccines gave positive results, confirming the presence of NDV. Multiple sequence alignment and phylogenetic analyses revealed that two of the vaccines had the lentogenic pathotype, while the other two had the mesogenic or velogenic pathotype.

Conclusion: This study provides information to facilitate strategies for regular control of the quality of vaccines in Nigeria.

Keywords: Newcastle disease virus, Nigeria, polymerase chain reaction, vaccines.

Impact of serum C-reactive protein level as a biomarker of cancer dissemination in canine lymphoid neoplasia

Research (Published online: 10-12-2022)

7. Impact of serum C-reactive protein level as a biomarker of cancer dissemination in canine lymphoid neoplasia

Nawin Manachai, Duangchanok Umnuayyonvaree, Panitnan Punyathi, Anudep Rungsipipat, and Kasem Rattanapinyopituk

Veterinary World, 15(12): 2810-2815

ABSTRACT

Background and Aim: C-reactive protein (CRP) is a highly sensitive but non-specific acute phase protein that has been widely used to predict the biological behavior of patients with cancer. This study aimed to examine the significance of the serum CRP biomarker in predicting the prognosis of dogs with lymphoma.

Materials and Methods: Blood samples (5 mL) were collected from 34 lymphoma dogs and control healthy dogs. Canine lymphoma clinical staging was classified using the World Health Organization (WHO) criteria. All lymphoma dogs were reclassified into two groups based on the disease stage. Stages IV and V were designated as advanced stages, and Stages I–III were designated as other stages. The serum CRP level was then determined using a commercial canine CRP fluorescent immunoassay kit and routine hematological and biochemical analyses. C-reactive protein levels, circulating inflammatory parameters, such as neutrophil-to-lymphocyte ratio, lymphocyte-to-monocyte ratio, and platelet-to-lymphocyte ratio, and albumin levels were compared between advanced stages (IV and V) and Stages I to III using Mann–Whitney U tests. Receiver operating characteristic (ROC) curves were also generated to determine the cutoff value, diagnostic sensitivity, and specificity of the CRP level.

Results: A prospective study identified 34 dogs recently diagnosed with canine lymphoma. C-reactive protein levels were significantly higher in lymphoma dogs in advanced stages (IV and V) than in lymphoma dogs in Stages I–III. According to the ROC curve analysis, a CRP cutoff level of 54.1 mg/L indicates advanced-stage canine lymphoma, which can be used as a biomarker to predict cancer dissemination.

Conclusion: Serum CRP concentrations can assist clinical decision-making on the WHO stage in lymphoma dogs in clinical applications. The limitations of this study include a small number of lymphomas and no survival analysis.

Keywords: advanced stage lymphoma, biomarker, dogs, serum C-reactive protein.

Phylogenetic analysis and antibiotic resistance of Escherichia coli isolated from wild and domestic animals at an agricultural land interface area of Salaphra wildlife sanctuary, Thailand

Research (Published online: 08-12-2022)

6. Phylogenetic analysis and antibiotic resistance of Escherichia coli isolated from wild and domestic animals at an agricultural land interface area of Salaphra wildlife sanctuary, Thailand

Taksaon Duangurai, Amporn Rungruengkitkul, Thida Kong-Ngoen, Witawat Tunyong, Nathamon Kosoltanapiwat, Poom Adisakwattana, Muthita Vanaporn, Nitaya Indrawattana, and Pornpan Pumirat

Veterinary World, 15(12): 2800-2809

ABSTRACT

Background and Aim: Domestic and wild animals are important reservoirs for antibiotic-resistant bacteria. This study aimed to isolate Escherichia coli from feces of domestic and wild animals at an agricultural land interface area of Salaphra Wildlife Sanctuary, Thailand, and study the phylogenic characteristics and antibiotic resistance in these isolates.

Materials and Methods: In this cross-sectional, descriptive study, we randomly collected ground feces from free-ranging wild animals (deer and elephants) and domestic animals (cattle and goats). All fecal samples were inoculated onto MacConkey agar plates, and lactose-fermenting colonies were identified as E. coli. Antibiotic susceptibility of the E. coli isolates was determined using the disc diffusion method. Polymerase chain reaction assays were used to detect antibiotic resistance and virulence genes.

Results: We obtained 362 E. coli isolates from the collected fecal samples. The E. coli isolates were categorized into four phylogenetic groups according to the virulence genes (chuA, vjaA, and TspE4C2). Phylogenetic Group D was predominant in the deer (41.67%) and elephants (63.29%), whereas phylogenetic Group B1 was predominant in the cattle (62.31%), and phylogenetic Groups A (36.36%) and B2 (33.33%) were predominant in the goats. Antibiotic susceptibility testing revealed that most antibiotic-resistant E. coli were isolated from domestic goats (96.96%). Among the 362 E. coli isolates, 38 (10.5%) were resistant to at least one antibiotic, 21 (5.8%) were resistant to two antibiotics, and 6 (1.66%) were resistant to three or more antibiotics. Ampicillin (AMP) was the most common antibiotic (48.48%) to which the E. coli were resistant, followed by tetracycline (TET) (45.45%) and trimethoprim-sulfamethoxazole (3.03%). One isolate from an elephant was resistant to five antibiotics: AMP, amoxicillin, sulfisoxazole, TET, and ciprofloxacin. Determination of antibiotic resistance genes confirmed that E. coli isolates carried antibiotic resistance genes associated with phenotypic resistance to antibiotics. Most antibiotic-resistant E. coli belonged to phylogenic Groups A and B1, and most non-resistant E. coli belonged to phylogenic Groups B2 and D.

Conclusion: Monitoring E. coli isolates from wild and domestic animals showed that all four phylogenic groups of E. coli have developed antibiotic resistance and are potential sources of multidrug resistance. High levels of antibiotic resistance have been linked to domestic animals. Our results support strengthening surveillance to monitor the emergence and effects of antibiotic-resistant microorganisms in animals.

Keywords: antibiotic resistance, domestic animal, Escherichia coli, phylogenic groups, phylogeny, wild animal.

Hematology and toll-like receptors 2 and 4 and lipopolysaccharide-induced tumor necrosis factor-α factor gene expression in peripheral blood mononuclear cells of Thai indigenous chickens

Research (Published online: 07-12-2022)

5. Hematology and toll-like receptors 2 and 4 and lipopolysaccharide-induced tumor necrosis factor-α factor gene expression in peripheral blood mononuclear cells of Thai indigenous chickens

Chananphat Tantikositruj, Asep Gunawan, Muhammad Jasim Uddin, Wirawan Nuchchanart, Chaiwat Boonkaewwan, Watchara Laenoi, and Autchara Kayan

Veterinary World, 15(12): 2795-2799

ABSTRACT

Background and Aim: Toll-like receptors (TLRs) play crucial roles in the early phase of infection in the innate immune response against bacteria, viruses, fungi, and parasites. Lipopolysaccharide-induced tumor necrosis factor-α factor (LITAF) is an essential transcription factor that regulates the immune system, apoptosis, and inflammatory cytokines. This study aimed to determine the hematological profile reflecting the immune response related to TLR2 and TLR4 and LITAF gene expression in Thai indigenous chickens.

Materials and Methods: Blood samples (2 mL) were randomly obtained from three chicken breeds (black-boned chicken, Fah Luang chicken, and Pradu Hang Dam chicken) at 16 weeks of age (n = 5 per breed). The hematological profile and mRNA expression within the peripheral blood mononuclear cells (PBMCs) were determined by hematological analysis and quantitative real-time polymerase chain reaction (qRT-PCR).

Results: The hematological profile differed significantly in terms of red blood cells (RBCs), hemoglobin, and white blood cells (WBCs) (p < 0.05). Black-boned chicken and Fah Luang chicken had lower RBC levels than Pradu Hang Dam chicken. Fah Luang chicken had lower hemoglobin than Pradu Hang Dam chicken. However, Fah Luang chicken had higher WBC levels than Pradu Hang Dam chicken. Hematocrit, heterophils, basophils, eosinophils, lymphocytes, and monocytes did not differ significantly among the groups (p > 0.05). According to qRT-PCR, the expression of the TLR2 gene did not differ significantly among the groups (p > 0.05), while TLR4 and LITAF gene expression did (p < 0.05). Toll-like receptor 4 and LITAF genes were most highly expressed in Fah Luang chicken.

Conclusion: The PBMCs of Thai indigenous chickens showed evidence of TLR4 and LITAF gene expression, with higher expression levels observed in Fah Luang chicken. From this preliminary study, it is concluded that TLR4 and LITAF genes might play roles in the main immune system response in Thai indigenous chickens.

Keywords: blood hematology, gene expression, immune, Thai indigenous chicken.

Multifarious feed additives on lamb performance on Kuwait farms

Research (Published online: 05-12-2022)

4. Multifarious feed additives on lamb performance on Kuwait farms

Hana'a Burezq and Faten Khalil

Veterinary World, 15(12): 2785-2794

ABSTRACT

Background and Aim: A change in the livestock feeding strategy is of utmost importance for the stability of animal health and sustainable livestock productivity to overcome the problem of subsiding the environmental effects of sheep production. Supplementing dietary feed with safe and efficient additives provides optimal animal performance and maximizes productivity. This study aimed to assess the effects of adding various feed additives to lamb rations for optimizing feed efficiency in weaned lambs for meat production in Kuwait.

Materials and Methods: The feed additives, namely, ammonium chloride, urea, algae, fishmeal, and humic acid, were investigated on the physical performance of lambs for their effect on body weight, length, height, and waist length. The total feed consumption rate and feed efficiency were also measured. Each treatment comprising five healthy lambs was randomly allocated into six treatments comprising 30 lambs. The six treatments were the basal ration supplemented with ammonium chloride (50–100 g/day/head), urea (30 g/day/head), fishmeal (35 g/day/head), algae (Spirulina platensis) powder (50 g/day/head), humic acid (2.5 g/day/head), control group with only basal ration. The study was conducted for around 27 months and the data were recorded once in 2 weeks.

Results: The results indicated a positive elevation in the physique of lambs with all tested additives, showing an affirmative insignia for lamb fattening. The growth parameters in terms of augmented length, height, and waist length of lambs' bodies amplified significantly with ammonium chloride and fishmeal supplement, while the other additives reported a non-significant increment. The feed consumption was significantly elevated for ammonium chloride, algae, and fishmeal supplementation, while humic acid was recorded the least. Concerning feed efficiency of young lambs, fish meal and ammonium chloride were reported best, followed by urea. In contrast, algae and humic acid exhibited a non-significant effect on feed efficiency.

Conclusion: This study exposed noteworthy influence on a lamb body's performance with the addition of fish meal and ammonium chloride in lamb rations, trailed by urea and algae.

Keywords: ammonium chloride, efficiency, feed additives, fishmeal, performance, urea.