The effect of Beauveria bassiana inoculation on plant growth, volatile constituents, and tick (Rhipicephalus appendiculatus) repellency of acetone extracts of Tulbaghia violacea

Research (Published online: 20-06-2020)

20. The effect of Beauveria bassiana inoculation on plant growth, volatile constituents, and tick (Rhipicephalus appendiculatus) repellency of acetone extracts of Tulbaghia violacea

Pumla Staffa, Nkululeko Nyangiwe, George Msalya, Yakob Petro Nagagi and Felix Nchu

Veterinary World, 13(6): 1159-1166

ABSTRACT

Aim: The purpose of the present study was to evaluate the effects of Beauveria bassiana (Hypocreales) inoculum on plant growth, volatile constituents, and tick repellency of the extracts of Tulbaghia violacea (Amaryllidaceae).

Materials and Methods: Eight-week-old potted seedlings of T. violacea were each inoculated with conidia of B. bassiana (strain SM3) suspended at a concentration of 1×106 conidia mL–1. Tissue colonization by fungal conidia was assessed after 3 weeks. Plant growth, volatile constituents, and tick repellency were assessed after 12 weeks post-treatment.

Results: B. bassiana conidia successfully colonized leaf and root tissues of T. violacea. The growth of fungal hyphae out of the leaf and root sections occurred in 75% and 91.6% of plants, respectively. Inoculation of the plants with B. bassiana significantly (p<0.05) influenced root length and plant height but did not have substantial effects on weights and leaf number of T. violacea. While the fungus did not have significant effects on overall number of the volatile chemical constituents, significant variations in the quantity (area ratio) were observed in at least four compounds that were detected. In the tick repellency bioassay, high concentration (20 w/v%) of acetone extract from fungus-exposed plants produced the least repellent effect on Rhipicephalus appendiculatus larvae (Ixodidae), while at lower concentrations (5 w/v% and 10 w/v%) of acetone extracts of T. violacea, tick repellent activity of the extract of the fungus treatment was significantly improved and was comparable to commercial N,N-Diethyl-m-toluamide and the other treatments.

Conclusion: Experimental fungal inoculation positively influenced plant growth in height and root length and tick (R. appendiculatus) repellency of acetone extracts of T. violacea at a concentration of 10 w/v% compared to the control treatment.

Keywords: Beauveria bassiana, fungal endophyte, secondary metabolites, tick repellency, tick toxicity, Tulbaghia violacea.

Detection of esophageal and glandular stomach calcification in cow (Bos taurus)

Research (Published online: 19-06-2020)

19. Detection of esophageal and glandular stomach calcification in cow (Bos taurus)

Esraa Abdul Khaliq Zegyer, Basim Abdullah Al Khuzaee and Ahmed Mahdi Al Badri

Veterinary World, 13(6): 1153-1158

ABSTRACT

Aim: The aims of this study were first to estimate calcification in the esophagus and abomasum of cows and second to quantify its appearance with increasing age using histopathological and immunohistochemical techniques.

Materials and Methods: Esophageal and abomasal samples from 24 healthy cows (Bos taurus) were collected. Hematoxylin and eosin, alizarin red, and von Kossa stains were used for histopathological analysis. Histopathological changes were confirmed with immunohistochemical staining, followed by digital image analysis.

Results: Histological findings revealed the esophagus and abomasum wall comprised four fundamental layers, the mucosa, submucosa, muscularis, and serosa or adventitia. At 1 year old, calcification was beginning to appear as fine diffused points in mucosa, submucosa, and muscularis of both esophagus and abomasum, appearing as small spots at 2 years old. With advancing age in all animals, this calcification began to appear as medium spots spread throughout all wall layers of these organs at 3 years old. By 4 years old, calcification had evolved into large dark foci spread substantially throughout the tunica submucosa and tunica muscularis. Immunohistochemical results exhibited positive immunoreaction to calcium salts in the esophagus and abomasum layers in all animals, which increased with age.

Conclusion: The current study concluded that calcification is a pathological event appearing spontaneously in various types of soft tissue, significantly increasing with age, either because of hypercalcemia and hyperphosphatemia or secondary to other diseases.

Keywords: abomasum, alizarin red, calcification, esophagus, immunohistochemistry, von Kossa.

Congestive heart failure in cattle; etiology, clinical, and ultrasonographic findings in 67 cases

Research (Published online: 19-06-2020)

18. Congestive heart failure in cattle; etiology, clinical, and ultrasonographic findings in 67 cases

Mustafa Abd El Raouf, Magdy Elgioushy and Shimaa A. Ezzeldein

Veterinary World, 13(6): 1145-1152

ABSTRACT

Background and Aim: Congestive heart failure (CHF) is a clinical disorder that results from cardiac dysfunction with subsequent fatal outcomes in most cases. Several diseases are incriminated in occurrence of CHF. Therefore, the aims of this study were to identify CHF etiology and associated clinical findings in 67 cows and to investigate the relationship between CHF and the other body organs using ultrasonographic examination.

Materials and Methods: Sixty-seven cows affected by CHF admitted to the clinic with a history of loss of appetite, decrease in milk production, constipation, and brisket edema were thoroughly investigated clinically and ultrasonographically. In addition, ten apparently healthy cows were used as a control group.

Results: Clinically, cows with CHF manifested jugular engorgement and pulsation (88.1%), brisket and/or intermandibular edema (77.6%), and muffled heart sounds (76.1%). Based on the ultrasonographic examination, traumatic pericarditis (82.1%) was the most prevalent etiology of CHF. Extracardiac etiology of CHF identified were exudative pleurisy (10.4%) and mediastinal abscesses (7.5%). Hepatomegaly (88.1%) and pleural effusion (61.2%) were the most documented consequences.

Conclusion: Both cardiac and extracardiac diseases could be associated with CHF in cattle. Ultrasonographic changes in liver and pleura secondary to CHF were the most common findings. Ultrasonography is a good tool for the diagnosis of cardiac and extracardiac etiologies of CHF in cattle.

Keywords: cattle, congestive heart failure, mediastinal abscess, pericarditis, pleurisy.

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.