ISMap02 element targeted nested polymerase chain in the detection of Mycobacterium avium subsp. paratuberculosis in fecal samples of cattle and buffaloes

Research (Published online: 31-03-2018)

22. ISMap02 element targeted nested polymerase chain in the detection of Mycobacterium avium subsp. paratuberculosis in fecal samples of cattle and buffaloes

Mamta Rani, Deepti Narang, Dinesh Kumar, Mudit Chandra, Sikh Tejinder Singh and G. Filia

Veterinary World, 11(3): 397-401

ABSTRACT

Background and Aim: Johne's disease is chronic granulomatous enteritis which affects ruminants. There are many diagnostic approaches for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) of which molecular detection methods using various elements are less time consuming and more accurate. The present study was conducted using ISMap02 element for nested polymerase chain reaction (nPCR) based detection of MAP in fecal samples. The aim was to test the sensitivity and specificity of the ISMap02 element and also to use this element for the detection of MAP in fecal samples of cattle and buffaloes.

Materials and Methods: A total of 211 fecal samples of cattle and buffaloes from different herds around Ludhiana aged between 2 and 13 years were collected, and DNA extraction was done from these samples. The nPCR was carried out for the detection of MAP in fecal samples.

Results: The ISMap02 element was specific for the detection of MAP only and showed a sensitivity of detection of 7.6 fg/μL of the standard genomic DNA. Among the 211 fecal samples of cattle and buffaloes tested for the ISMap02 element, 18 samples (8.5%) were positive for MAP.

Conclusion: The ISMap02 element is specific and sensitive for the detection of MAP in various samples, and when used in nPCR format, it can increase the sensitivity of detection.

Keywords: ISMap02, Mycobacterium avium subsp. paratuberculosis, nested polymerase chain reaction, paratuberculosis.

Inventory of lice of mammals and farmyard chicken in North-eastern Algeria

Research (Published online: 30-03-2018)

21. Inventory of lice of mammals and farmyard chicken in North-eastern Algeria

Mohamed Nadir Meguini, Souad Righi, Faycal Zeroual, Khelaf Saidani and Ahmed Benakhla

Veterinary World, 11(3): 386-396

ABSTRACT

Background and Aim: Lice are permanent ectoparasites, extremely specific to their hosts. Their great importance in veterinary medicine remain significant, they can cause their direct pathogenic actions like irritability, dermatitis, anemia, decreased weight gain, and milk production. The purpose of this work was to made the first time an inventory of mammalian lice in North-eastern Algeria.

Materials and Methods: Our survey of lice infestation was conducted on several animal species from five provinces of North-eastern Algeria. A total of 57 cattle, 83 sheep, 77 goats, 111 wild boars, and 63 farmyard chickens were examined. The collection of lice was carried out much more in mammals and chickens during the winter period. Lice were collected either manually or using brushing and kept in flasks containing 70% ethanol. The identification of lice was achieved in the laboratory using a binocular loupe.

Results: Concerning cattle, 63% and 27% of those examined subjects from Souk-Ahras and Guelma study areas, respectively, were carriers of lice. Damalinia bovis was the louse most frequently found on cattle in these two regions. Three other species were identified in Souk-Ahras: Haematopinus eurysternus (25%), Linognathus vituli (10%), and Solenopotes capillatus (5%). Regarding sheep, 39% and 24% of examined animals in Souk-Ahras and Guelma, were carrying lice. Damalinia ovis was the most frequently encountered lice on sheep in both regions. Linognathus ovillus also was identified in Souk-Ahras, representing 0.3% of the collected lice. Concerning goats, 53% and 30% of examined animals in Souk-Ahras and Guelma, were parasitized of lice. Two species of lice were found: Damalinia caprae and Linognathus africanus. For farmyard chickens, 69% and 100% of the farmyard chicken in Souk-Ahras and Mila were parasitized by lice, respectively. Menopon gallinae was the most frequently encountered louse in farmyard chicken in both regions. Eight other species were identified in Mila and four other species only in Souk-Ahras. Finally, 25% and 28% of the wild boars in Annaba and El Tarf were parasitized by lice, respectively. Haematopinus suis was the only species found on wild boars in both regions.

Conclusion: These results are to be taken into account for lice control schemes and louse-borne diseases.

Keywords: boars, farmyard chickens, lice, North-eastern Algeria, ruminants.

Isolation of Shiga toxin-producing Escherichia coli harboring variant Shiga toxin genes from seafood

Research (Published online: 28-03-2018)

20. Isolation of Shiga toxin-producing Escherichia coli harboring variant Shiga toxin genes from seafood

Sreepriya Prakasan, Parmanand Prabhakar, Manjusha Lekshmi, Binaya Bhusan Nayak and Sanath Kumar

Veterinary World, 11(3): 379-385

ABSTRACT

Background and Aim: Shiga toxin-producing Escherichia coli (STEC) are important pathogens of global significance. STEC are responsible for numerous food-borne outbreaks worldwide and their presence in food is a potential health hazard. The objective of the present study was to determine the incidence of STEC in fresh seafood in Mumbai, India, and to characterize STEC with respect to their virulence determinants.

Materials and Methods: A total of 368 E. coli were isolated from 39 fresh seafood samples (18 finfish and 21 shellfish) using culture-based methods. The isolates were screened by polymerase chain reaction (PCR) for the genes commonly associated with STEC. The variant Shiga toxin genes were confirmed by Southern blotting and hybridization followed by DNA sequencing.

Results: One or more Shiga toxins genes were detected in 61 isolates. Of 39 samples analyzed, 10 (25.64%) samples harbored STEC. Other virulence genes, namely, eaeA (coding for an intimin) and hlyA (hemolysin A) were detected in 43 and 15 seafood isolates, respectively. The variant stx1 genes from 6 isolates were sequenced, five of which were found to be stx1d variants, while one sequence varied considerably from known stx1sequences. Southern hybridization and DNA sequence analysis suggested putative Shiga toxin variant genes (stx2) in at least 3 other isolates.

Conclusion: The results of this study showed the occurrence of STEC in seafood harboring one or more Shiga toxin genes. The detection of STEC by PCR may be hampered due to the presence of variant genes such as the stx1d in STEC. This is the first report of stx1d gene in STEC isolated from Indian seafood.

Keywords: Escherichia coli, pathogen, seafood, Shiga toxin, Shiga toxin-producing Escherichia coli, virulence gene.

Antibacterial and antioxidant activity of Juniperus thurifera L. leaf extracts growing in East of Algeria

Research (Published online: 27-03-2018)

19. Antibacterial and antioxidant activity of Juniperus thurifera L. leaf extracts growing in East of Algeria

Merradi Manel, Heleili Nouzha, Mekari Rim, Mekkaoui Imane, Aouachria Sana, Oucheriah Yasmine and Ayachi Ammar

Veterinary World, 11(3): 373-378

ABSTRACT

Aim: This work aimed to evaluate the biological activity of the leaf extracts of Juniperus thurifera L., which is an Algerian endemic tree that belongs to the family of Cupressaceae.

Materials and Methods: The plant leaves were extracted in solvents of increasing polarity to obtain different extracts such as methanol, petroleum ether, chloroform, ethyl acetate, and aqueous extracts (MeE, PEE, ChlE, EtAE, and AqE). The antioxidant activity of four extracts (MeE, ChlE, EtAE, and AqE) was assessed by trapping test of 1,1-diphenyl-2- picrylhydrazyl (DPPH) radical. The evaluation of antibacterial activity of MeE, ChlE, EtAE, and PEE was done using the disk diffusion method on solid agar.

Results: The three extracts of EtAE, AqE, and MeE showed high antiradical activity toward the DPPH radical (IC50=29.348 μg/mL, 37.538 μg/mL, and 52.573 μg/mL, respectively), while the lowest radical scavenging activity was expressed by the ChlE (IC50=70.096 μg/mL). These extracts were active only toward the Gram-positive bacteria (Staphylococcus aureus ATCC and methicillin-resistant S. aureus) at different concentrations, and the highest activity was obtained with the ChlE with an inhibition diameter of 14 mm at the concentration of 1 g/mL. No inhibition was detected for all of these extracts against the Gram-negative tested strains (Escherichia coli ATCC, Pseudomonas aeruginosa ATCC, and Enterobacter cloacae (extended spectrum β-lactamase).

Conclusion: From this study, on the one hand, it was concluded that J. thurifera L. leaves extracts exhibited a very intense antioxidant potential toward the DPPH radical, and on the other hand, the antibacterial activity showed an action spectrum exclusively toward the Gram-positive bacteria.

Keywords: antibacterial activity, antioxidant activity, extraction, Juniperus thurifera L., trapping test of 1,1-diphenyl-2- picrylhydrazyl radical.

Evaluation of antimycobacterial activity of Curcuma xanthorrhiza ethanolic extract against Mycobacterium tuberculosis H37Rv in vitro

Research (Published online: 27-03-2018)

18. Evaluation of antimycobacterial activity of Curcuma xanthorrhiza ethanolic extract against Mycobacterium tuberculosis H37Rv in vitro

Ngadino, Setiawan, Koerniasari, Ernawati and S. A. Sudjarwo

Veterinary World, 11(3): 368-372

ABSTRACT

Aim: The aim of this study was to evaluate the antimycobacterial activity of the Curcuma xanthorrhiza ethanolic extract in vitro.

Materials and Methods: Ethanolic extract of C. xanthorrhiza was set by maceration method. The broth microdilution and disc diffusion method were used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC), respectively, of C. xanthorrhiza ethanol extract on strain Mycobacterium tuberculosis H37Rv.

Results: C. xanthorrhiza ethanol extract was found to have the antimycobacterial effects with a MIC value of 1600 μg/ml while MBC value of 3200 μg/ml for M. tuberculosis H37Rv.

Conclusion: From these findings , it can be concluded that C. xanthorrhiza ethanol extract have an antibacterial activity against Mycobacterium tuberculosis H37Rv in vitro and its potency elevated by increasing the C. xanthorrhiza ethanol extract concentration.

Keywords: antimycobacterial, Curcuma xanthorrhiza, minimal bactericidal concentration, minimal inhibitory concentration.

Detection of Brucella spp. in milk from seronegative cows by real-time polymerase chain reaction in the region of Batna, Algeria

Research (Published online: 26-03-2018)

17. Detection of Brucella spp. in milk from seronegative cows by real-time polymerase chain reaction in the region of Batna, Algeria

Rabehi Sabrina, Hamdi Taha Mossadak, Mamache Bakir, Meghezzi Asma and Boushaba Khaoula

Veterinary World, 11(3): 363-367

ABSTRACT

Aim: The aim of this study was to detect Brucella spp. DNA in milk samples collected from seronegative cows using the real-time polymerase chain reaction (PCR) assay for diagnosis of brucellosis in seronegative dairy cows to prevent transmission of disease to humans and to reduce economic losses in animal production.

Materials and Methods: In this study, 65 milk samples were investigated for the detection of Brucella spp. The detection of the IS711 gene in all samples was done by real-time PCR assay by comparative cycle threshold method.

Results: The results show that of the 65 DNA samples tested, 2 (3.08%) were positive for Brucella infection. The mean cyclic threshold values of IS711 real-time PCR test were 37.97 and 40.48, indicating a positive reaction.

Conclusion: The results of the present study indicated that the real-time PCR appears to offer several advantages over serological tests. For this reason, the real-time PCR should be validated on representative numbers of Brucella-infected and free samples before being implemented in routine diagnosis in human and animal brucellosis for controlling this disease.

Keywords: Brucella spp., milk, real-time polymerase chain reaction, seronegative cows.

Isolation and identification of bacteria causing mastitis in small ruminants and their susceptibility to antibiotics, honey, essential oils, and plant extracts

Research (Published online: 26-03-2018)

16. Isolation and identification of bacteria causing mastitis in small ruminants and their susceptibility to antibiotics, honey, essential oils, and plant extracts

Abeer Mostafa Abdalhamed, Gamil Sayed Gamil Zeedan and Hala Abdoula Ahmed Abou Zeina

Veterinary World, 11(3): 355-362

ABSTRACT

Aim: The present work aims to isolate and identify bacteria that cause mastitis in small ruminants and evaluates the antibacterial activity of some antibiotics, honey, essential oils, and plant extracts.

Materials and Methods: A total of 289 milk samples were collected from udder secretions of sheep (n=189) and goat (n=100) from El-Fayoum, Beni-Suef, and Giza governorates. Screening subclinical mastitis (SCM) was done using California Mastitis Test (CMT); identification of the isolates was achieved using Gram's staining, hemolytic pattern, colony morphology, and biochemical tests using Analytical Profile Index.

Results: On clinical examination, the incidence of clinical mastitis (CM) was found to be 5.88% and 7% in sheep and goat, respectively. On CMT, SCM was found to be 25 (13.23%) and 11 (10%) in sheep and goat, respectively. Bacteriological examination of all milk samples found the presence of Staphylococcus aureus (SA) (31.1%), coagulase-negative staphylococci (CNS) (19.5%), Escherichia coli (EC) (8.3%), Streptococcus spp. (5.6%), Klebsiella spp. (3.77%), and Pseudomonas spp. (1.89%), while no bacteria were cultured from 81.66% of the samples. Identification of 9 isolates of CNS was achieved by using API staph test to Staphylococcus epidermidis, Staphylococcus hominis, Staphylococcus cohnii, and Staphylococcus saprophyticus. The highest bacterial resistance was found in EC (67.14%) followed by Kp (45.28%) and SA (26.57%).

Conclusion: Onion and black cumin essential oils followed by Egyptian honey showed strong antibacterial effects against multidrug-resistant bacteria. Finally, our study proved that Egyptian honey, onion, and black cumin essential oils have a marked strong antibacterial effect against bacteria isolated from small ruminant mastitis, but still further extensive studies are needed to discover the therapeutic properties of these plant extracts and honey.

Keywords: black cumin, essential oil, honey, mastitis, onion, small ruminants, Staphylococcus aureus.