Determining highly pathogenic H5 avian influenza clade 2.3.2.1c seroprevalence in ducks, Purbalingga, Central Java, Indonesia

Research (Published online: 18-06-2020)

17. Determining highly pathogenic H5 avian influenza clade 2.3.2.1c seroprevalence in ducks, Purbalingga, Central Java, Indonesia

Imas Yuyun, Hendra Wibawa, Gunawan Setiaji, Tri Anggraeni Kusumastuti, and Widagdo Sri Nugroho

Veterinary World, 13(6): 1138-1144

ABSTRACT

Background and Aim: In Indonesia, highly pathogenic avian influenza (HPAI) H5N1 outbreaks in poultry are still reported. The disease causes a decrease in egg production and an increase in mortality; this has an impact on the economic losses of farmers. Several studies have considered that ducks play a role in the HPAI endemicity in the country; however, little is known about whether or not the type of duck farming is associated with HPAI H5 virus infection, particularly within clade 2.3.2.1c, which has been predominantly found in poultry since 2014. A cross-sectional study was conducted to determine the HPAI seroprevalence for H5 subtype clade 2.3.2.1c in laying ducks that are kept intensively and nomadically and to determine the associated risk factors.

Materials and Methods: Forty-nine duck farmers were randomly selected from ten sub-districts in Purbalingga District, Central Java, Indonesia; a cross-sectional study was implemented to collect field data. Based on an expected HPAI prevalence level of 10%, estimated accuracy of ± 5%, and 95% confidence interval (CI), the total sample size was calculated at 36 individuals. Samples must be multiplied by 7 to reduce bias; thus, 252 ducks were taken as samples in this study. Considering that the maintenance and duck handling were uniform and farmers complained that the effect of activity to take duck samples would reduce egg production, this study only took samples from 245 ducks (oropharyngeal swabs and serum). Those samples were taken from five birds on each farm. Hemagglutination inhibition tests examined the serum samples for HPAI H5 Clade 2.3.2.1c, and pool swab samples (five swabs in one viral media transport) were examined by real-time reverse transcription-polymerase chain reaction (qRT-PCR) test for influenza Type A and H5 subtype virus. Information regarding farm management was obtained using a questionnaire; face-to-face interviews were conducted with the duck farmers using native Javanese language.

Results: Serum and swabs from 245 ducks were collected in total. For individual birds, 54.69% (134/245) of serum samples were H5 seropositive. Seroprevalence among nomadic ducks was 59.28% (95% CI: 0.48-0.61), which was higher than among intensively farmed ducks (48.57%, 95% CI: 0.38-0.58). Farm-level seroprevalence was 50% (95% CI: 0.30-0.69) for nomadic ducks but only 28.57% (95% CI: 0.11-0.51) for intensively farmed ducks. The farm-level virus prevalence (proportion of flocks with at least one bird positive for influenza Type A) was 17.85% (95% CI: 0.07-0.35) for nomadic ducks and 4.76% (1/21) for intensively farmed ducks (95% CI: 0.008-0.23). All influenza Type A positive samples were negative for the H5 subtype, indicating that another HA subtype AI viruses might have been circulating in ducks in the study area. A relationship between duck farms that were H5 seropositive and their maintenance system was present; however, this relationship was not significant, the nomadic duck system detected 2 times higher H5-seropositive ducks than the intensive farming system (OR: 2.16, 95% CI: 0.33-14.31).

Conclusion: This study found that the seroprevalence of HPAI in the duck population level in Purbalingga was 54.69% and demonstrated that the nomadic duck farming system was more likely to acquire HPAI H5 infection than the intensive farming duck system. Other risk factors should be further investigated as the diversity of the farming system is partially related to HPAI H5 infection.

Keywords: avian influenza, ducks, farming system, highly pathogenic avian influenza H5, risk factors, seroprevalence.

Evaluation of two fixation techniques for direct observation of biofilm formation of Bacillus subtilis in situ, on Congo red agar, using scanning electron microscopy

Research (Published online: 18-06-2020)

16. Evaluation of two fixation techniques for direct observation of biofilm formation of Bacillus subtilis in situ, on Congo red agar, using scanning electron microscopy

Nadia Mahmoud Tawfiq Jebril

Veterinary World, 13(6): 1133-1137

ABSTRACT

Background and Aim: Direct observation, scanning electron microscopy (SEM) is a common method used for the observations of biofilms. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide) (EDC) fixation method has proven to be a valuable fixation method in the observation of these biofilms. Still, it entails a method of biofilm fixation that can damage slim structures, leading to the impossible observation of biofilm development. In contrast, alcian blue and lysine (ABL) fixation technique appears more glycocalyx of biofilm, fully preserved samples, which may provide much insight into the development of B. subtilis biofilms.

Materials and Methods: Here, the evaluation of the fixation of ABL technique for the study of B. subtilis biofilms was carried out in situ, on Congo red agar. In doing so, the comparison to commonly use conventional EDC technique for sample fixation, and observation was carried out. Observations were based on SEM over 30 samples.

Results: Overall, ABL technique provided excellent observation of biofilms formed in situ, on Congo red agar, and revealed slime structures, which have not been observed, much in standard EDC fixation or earlier in other studies of these biofilms in B. subtilis.

Conclusion: This study reported the appropriate use of ABL in the fixation technique for the preservation of biofilm of B. subtilis.

Keywords: alcian blue, Bacillus subtilis, biofilm, Congo red agar, scanning electron microscopy.

Single-nucleotide polymorphisms in FABP4 gene associated with growth traits in Egyptian sheep

Research (Published online: 17-06-2020)

15. Single-nucleotide polymorphisms in FABP4 gene associated with growth traits in Egyptian sheep

Heba Ibrahim Shafey, Karima Fathy Mahrous, Amal Ahmed Mohamed Hassan, Hossam Eldin Rushdi and Mohamed Abd El-Aziz Mohamed Ibrahim

Veterinary World, 13(6): 1126-1132

ABSTRACT

Aim: The present study was performed to assess the association of single-nucleotide polymorphisms (SNPs) in the fatty acid-binding protein 4 (FABP4) gene with birth weight (BW), final weight (FW), and average daily gain (ADG) in three Egyptian sheep breeds.

Materials and Methods: Genomic DNA was extracted from the blood samples of 50 male and female individuals representing Ossimi, Rahmani, and Barki sheep breeds. A 407 bp nucleotide (nt) segment from the first intron of FABP4 was amplified by polymerase chain reaction, sequenced, and analyzed in the different samples.

Results: Sequence analysis of the determined segment (407 bp) revealed four SNPs (all transition types) at nt position 372 (CP011894.1:g.57605471) A>G, nt position 211 (CP011894.1:g.57605632) A>G, nt position 143 (CP011894.1:g.57605700) T>C, and nt position 111 (CP011894.1:g.57605732) T>C. The allelic and genotypic frequencies for the identified SNPs in the sheep breeds were calculated. At nt positions 372 and 211, two alleles were identified (A and G). Only two genotypes were present at nt position 372 (AA and AG), while three genotypes were present at nt position 211 (AA, AG, and GG). Two alleles (T and C) and three identified genotypes (TT, TC, and CC) were detected at nt positions 143 and 111. Analysis of the results revealed that AA genotype at nt position 372 is associated with higher estimates for BW, FW, and ADG when compared to all the other genotypes. Very high correlation coefficients were found between the genotypes 143-TT and 111-TT and also between 143-TC and 111-TC. The genotypes 372-AG, 211-GG, 211-AA, 143-TT, 143-CC, 111-TT, 111-TC, and 111-CC were associated with negative effects on BW, FW, and ADG.

Conclusion: The detection of four SNPs in a partial sequence of the Egyptian ovine FABP4 gene intron 1 reflected that this gene harbors substantial diversity. In addition, a novel SNP at nt position 372 (CP011894.1:g.57605471) A>G was associated with higher estimates for BW, FW, and ADG.

Keywords: body weight, FABP4 gene, growth rate, sheep, single-nucleotide polymorphisms.

Differential DNA methylation analysis across the promoter regions using methylated DNA immunoprecipitation sequencing profiling of porcine loin muscle

Research (Published online: 16-06-2020)

14. Differential DNA methylation analysis across the promoter regions using methylated DNA immunoprecipitation sequencing profiling of porcine loin muscle

Kaj Chokeshaiusaha, Denis Puthier, Thanida Sananmuang, Em-on Olanratmanee, Catherine Nguyen and Roongtham Kedkovid

Veterinary World, 13(6): 1113-1125

ABSTRACT

Background and Aim: Pork leanness and marbling are among the essential traits of consumer preference. To acquire knowledge about universal epigenetic regulations for improving breed selection, a meta-analysis of methylated DNA immunoprecipitation sequencing (MeDIP-seq) profiling data of mixed loin muscle types was performed in this study.

Materials and Methods: MeDIP-seq profiling datasets of longissimus dorsi muscle and psoas major muscles from male and female pigs of Landrace and Tibetan breeds were preprocessed and aligned to the porcine genome. Analysis of differential methylated DNA regions (DMRs) between the breeds was performed by focusing on transcription start sites (TSSs) of known genes (–20,000-3000 bases from TSS). All associated genes were further reviewed for their functions and predicted for transcription factors (TF) possibly associated with their TSSs.

Results: When the methylation levels of DMRs in TSS regions of Landrace breed were compared to those of Tibetan breed, 10 DMRs were hypomethylated (Landrace < Tibetan), and 19 DMRs were hypermethylated (Landrace > Tibetan), accordingly (p≤0.001). According to the reviews about gene functions, all associated genes were pieces of evidence for their roles in a variety of muscle and lipid metabolisms. Prediction of the binding TFs revealed the six most abundant binding TFs to such DMRs-associated TSS (p≤0.0001) as follows: ZNF384, Foxd3, IRF1, KLF9, EWSR1-FLI1, HES5, and TFAP2A.

Conclusion: Common DMRs-associated TSS between the lean-type and the marbled-type loin muscles were identified in this study. Interestingly, the genes associated with such regions were strongly evidenced for their possible roles on the muscle trait characteristics by which further novel research topics could be focused on them in the future.

Keywords: differential DNA methylation, lean, loin muscle, marbled, methylated DNA immunoprecipitation sequencing, transcription start site.

Detection of feline idiopathic cystitis as the cause of feline lower urinary tract disease in Sleman Regency, Indonesia

Research (Published online: 16-06-2020)

13. Detection of feline idiopathic cystitis as the cause of feline lower urinary tract disease in Sleman Regency, Indonesia

Andi Tri Julyana Eka Astuty, Ida Tjahajati and Widagdo Sri Nugroho

Veterinary World, 13(6): 1108-1112

ABSTRACT

Background and Aim: Feline lower urinary tract disease (FLUTD) is one of the common cat diseases. The aim of this study was to detect feline idiopathic cystitis (FIC) as a cause of FLUTD in Sleman Regency, which is a problem in the population.

Materials and Methods: Seventy-three cats with FLUTD symptoms were used from seven veterinary practices in Sleman Regency. The logging of each cat's medical history, clinical examination, urinalysis, routine blood screening, and ultrasonography was conducted to diagnose the cause of FLUTD.

Results: The percentages of diseases causing FLUTD included FIC 21.9%, urolithiasis 57.5%, urinary tract infection (UTI) 16.4%, neoplasia 1.4%, trauma 1.4%, and nervous disorders 1.4%.

Conclusion: FIC, one of the causes of FLUTD, is found in cats and has become a problem among the cat population in Sleman Regency, Yogyakarta, Indonesia. Various handling and preventive efforts should be undertaken against the disease.

Keywords: cat, detection, feline idiopathic cystitis, feline lower urinary tract disease, Indonesia, Sleman.

Genotyping of rabbit hemorrhagic disease virus isolated from diseased rabbits in Egyptian Provinces by VP 60 sequencing

Research (Published online: 15-06-2020)

12. Genotyping of rabbit hemorrhagic disease virus detected in diseased rabbits in Egyptian Provinces by VP60 sequencing

Ahmed M. Erfan and Azhar G. Shalaby

Veterinary World, 13(6): 1098-1107

ABSTRACT

Background and Aim: Rabbit hemorrhagic disease (RHD) is an economically important disorder of rabbits, where infection results in severe losses to the meat and fur industries. Our goal was to characterize the RHD virus (RHDV) strains currently circulating in different regions of Egypt.

Materials and Methods: Fifty rabbits suspected of harboring RHDV from 15 Egyptian governorates were evaluated. Diseased rabbits were identified by clinical signs and postmortem lesions. RHDV was confirmed through hemagglutination assay (HA) and polymerase chain reaction (PCR). Partial sequencing of the VP60 gene was performed for genotyping.

Results: From 50 rabbits, we identified 16 cases of RHDV (32%) by HA and PCR, including seven males and nine females. We identified two distinct genotypes through sequencing of an amplified fragment of the virus VP 60 gene. One group is composed of those circulating primarily in upper Egypt, which is closely related to the classical G3-G5 virus strains, and the second group, circulating predominantly in lower Egypt, was more closely related to the RHDV2 variant. The overall nucleotide sequence identity ranged from 78.4% to 100%, and identity with the vaccine strains ranged from 78.8% to 91.1%.

Conclusion: Our results constitute important documentation of RHDV strains currently circulating in Egypt. The findings suggest that there may be a limit to the effectiveness of currently applied vaccine strains as this formulation may not cover all circulating strains. A wider investigation that includes both domestic and wild rabbits will be needed to identify appropriate control measures for this disease.

Keywords: Egypt, native rabbits, prevalence, rabbit hemorrhagic disease virus, VP60.

The influence of "Stimix Zoostim" and "Normosil" probiotics on fecal microflora, hematologic indicators, nutrient digestibility, and growth of mother-bonded calves

Research (Published online: 15-06-2020)

11. The influence of "Stimix Zoostim" and "Normosil" probiotics on fecal microflora, hematologic indicators, nutrient digestibility, and growth of mother-bonded calves

Fail Khaziakhmetov, Airat Khabirov, Khamit Tagirov, Ruzil Avzalov, Gulnara Tsapalova and Almaz Basharov

Veterinary World, 13(6): 1091-1097

ABSTRACT

Aim: This paper presents the results of the studies on "Stimix Zoostim" and "Normosil" probiotics and their influence on fecal microflora, hematologic indicators, immunoresistance, nutrient digestibility, and growth intensity of mother-bonded calves.

Materials and Methods: The calves of the control group were fed with their basic diet (BD) without "Stimix Zoostim" or "Normosil", whereas the calves of the experimental group were fed with their BD supplemented with "Stimix Zoostim" and "Normosil". For 10-20-day-old calves, the daily dose was 10 mL per head, whereas 21-90-day-old calves received 15 mL of probiotics per head per day. The calves of the experimental group were administered probiotics every day. Calves aged 10 to 60 days received probiotics with milk, and then at the age of 61 to 90 days, received probiotics with water. (This is a sequential process. At 60 days old, calf rearing with milk stops, and after that, we use probiotics added to water to rear them). Both groups were administered probiotics twice a day, specifically 50% of the daily dose at each feeding time.

Results: The results in this study proved that "Stimix Zoostim" and "Normosil" probiotics exhibit high probiotic activity and have a positive effect on the calves' fecal microflora. Feeding the calves with probiotics resulted in a significant increase in the number of normal flora, such as lactic acid bacteria and bifidobacteria, and a decrease in the amount of Escherichia. It also resulted in an increase in red blood cells, hemoglobin, and ?-globulins within the physiological range. In our study, we found that the phagocytic reaction in the blood serum of the experimental group was slightly enhanced, suggesting a high response of the body to infectious agents.

Conclusion: Thus, the group of calves receiving probiotic "Stimix Zoostim" exhibited an exceedance of phagocytic activity by 4.8% and the group receiving "Normosil" by 4.4%, in comparison with the control group. The daily dose of 10 mL of probiotics per head for 10-20-day-old calves and 15 mL per head for 21-90-day-old calves also had a positive effect on nutrient digestibility, growth, and forage consumption. The economic benefit per animal was 149.23 and 157.0 rubles, respectively.

Keywords: average gain, blood immunoresistance, forage consumption, hematologic indicators, mother-bonded calves, nutrient digestibility.