Detection and characterization of pathogenic Pseudomonas aeruginosa from bovine subclinical mastitis in West Bengal, India

Research (Published online: 04-07-2017)

4. Detection and characterization of pathogenic Pseudomonas aeruginosa from bovine subclinical mastitis in West Bengal, India

S. Banerjee, K. Batabyal, S. N. Joardar, D. P. Isore, S. Dey, I. Samanta, T. K. Samanta and S. Murmu

Veterinary World, 10(7): 738-742

ABSTRACT

Aim: Subclinical mastitis in bovines is mainly responsible for the huge economic loss of the dairy farmers, of which Pseudomonas aeruginosa is one of the causative agents. The study was aimed at a screening of suspected milk samples from different cattle farms of West Bengal for detection and confirmation of P. aeruginosa strains followed by their characterization.

Materials and Methods: Around 422 milk samples were screened from different dairy farms primarily by on-spot bromothymol blue (BTB) test and then in the lab by somatic cell counts (SCC) to finally consider 352 samples for detection of P. aeruginosa. Selective isolation and confirmation of the isolates were done using selective media, viz., cetrimide and Pseudomonas agar followed by confirmation by fluorescent technique. Molecular characterization of the strains was done by polymerase chain reaction for the presence of toxA (enterotoxin A, 352 bp) and exoS (exoenzyme S, 504 bp) genes.

Results: Approximately, 371 (87.9%) samples were positive in on-spot BTB test among which 352 (94.8%) samples revealed high SCC values (more than 3 lakh cells/ml) showing infection when screened. Among these, 23 (6.5%) samples yielded typical Pseudomonas sp. isolates out of which only 19 (5.4%) isolates were confirmed to be P. aeruginosa which showed characteristic blue-green fluorescence due to the presence of pigment pyoverdin under ultraviolet light. Out of these 19 isolates, 11 isolates were positive for toxA, 6 isolates for exoS, and 2 for both these pathogenic genes.

Conclusion: Approximately, 5.4% cases of bovine subclinical mastitis infections in South Bengal were associated with P. aeruginosa which possess pathogenic genes such as toxA (63.2%) and exoS (36.8%).

Keywords: bovines, characterization, exoS, Pseudomonas aeruginosa, subclinical mastitis, toxA.

Cuticular surface damage of Ascaridia galli adult worms treated with Veitchia merrillii betel nuts extract in vitro

Research (Published online: 03-07-2017)

3. Cuticular surface damage of Ascaridia galli adult worms treated with Veitchia merrillii betel nuts extract in vitro

Ummu Balqis, Muhammad Hambal, Rinidar, Farida Athaillah, Ismail, Azhar, Henni Vanda and Darmawi

Veterinary World, 10(7): 732-737

ABSTRACT

Aim: The objective of this research was to in vitro evaluate the cuticular surface damage of Ascaridia galli adult worms treated with ethanolic extract of betel nuts Veitchia merrillii.

Materials and Methods: Phytochemical screening was done using FeCl3, Wagner and Dragendorff reagents, NaOH, MgHCl, and Liebermann-Burchard reaction test. Amount of 16 worms were segregated into four groups with three replicates. Four worms of each group submerged into phosphate buffered saline, 25 mg/ml, and 75 mg/ml crude ethanolic extract of V. merrillii, and 15 mg/ml albendazole. The effect of these extract was observed 40 h after incubation as soon as worms death. The worms were sectioned transversally and were explored for any cuticular histopathological changes in their body surface under microscope.

Results: We found that the ethanolic extract of V. merrillii betel nuts contains tannins, alkaloids, flavonoids, triterpenoids, and saponins. The ethanolic extract of betel nuts V. merrillii induces surface alterations caused cuticular damage of A. galli adult worms.

Conclusion: We concluded that ethanolic extract of betel nuts V. merrillii possess anthelmintic activity caused cuticular damage of A. galli adult worms.

Keywords: anthelmintics, Ascaridia galli, cuticle, Veitchia merrillii.

Performance of broiler chicken fed multicarbohydrases supplemented low energy diet

2. Performance of broiler chicken fed multicarbohydrases supplemented low energy diet

Kumar Govil, Sunil Nayak, R. P. S. Baghel, A. K. Patil, C. D. Malapure and Dinesh Thakur

Veterinary World, 10(7): 727-731

doi: 10.14202/vetworld.2017.727-731

Abstract

Aim: Objective of this study was to investigate the effect of multicarbohydrases supplementation on performance of broilers fed low energy diet.

Materials and Methods: A total of 75 days old chicks were selected and randomly divided into three treatments groups (T₁, T₂, and T₃); each group contained 25 chicks distributed in five replicates of five chicks each. T₁ group (positive control) was offered control ration formulated as per Bureau of Indian Standards recommendations. In T₂ group (negative control) ration, metabolizable energy (ME) was reduced by 100 kcal/kg diet. T₃ group ration was same as that of T₂ except that it was supplemented with multicarbohydrases (xylanase at 50 g/ton+mannanase at 50 g/ton+amylase at 40 g/ton). Feed intake and body weight of all experimental birds were recorded weekly. Metabolic trial was conducted for 3 days at the end of experiment to know the retention of nutrients.

Results: Significant improvement (p<0.01) was observed in total weight gain, feed conversion efficiency, and performance index in broilers under supplementary group T₃ as compared to T₁ and T₂ groups. Retention of crude protein and ether extract was significantly increased (p<0.05) in T₃ group supplemented with multicarbohydrases as compared to other groups. Retention of dry matter, crude fiber, and nitrogen-free extract was comparable in all the three groups. Significantly highest dressed weight, eviscerated weight, and drawn weight (% of live body weight) were observed in multicarbohydrases supplemented T₃ group, however it was comparable in T₁ and T₂ groups.

Conclusion: It was concluded that the supplementation of multicarbohydrases (xylanase at 50 g/ton+mannanase at 50 g/ton+amylase at 40 g/ton) in low energy diet improved overall performance of broilers.

Keywords: broiler, carcass traits, multicarbohydrases, performance.

Screening method for Staphylococcus aureus identification in subclinical bovine mastitis from dairy farms

Research (Published online: 01-07-2017)

1. Screening method for Staphylococcus aureus identification in subclinical bovine mastitis 

from dairy farms

Natapol Pumipuntu, Suphang Kulpeanprasit, Sirijan Santajit, Witawat Tunyong, Thida Kong-ngoen, 

Woranich Hinthong and Nitaya Indrawattana

Veterinary World, 10(7): 721-726

ABSTRACT

Background: Staphylococcus aureus is one of the most important contagious bacteria causing subclinical bovine mastitis. This bacterial infection is commonly identified by determine the pathogen in bovine milk samples through conventional technique including coagulase test. However, this test has several disadvantages as low sensitivity, risk of biohazard, cost expensive, and limited preparation especially in local area.

Aim: Aim of this study was to compare and assess the screening method, Mannitol fermentation test (Mannitol salt agar [MSA]), and deoxyribonuclease (DNase) test, for S. aureus identification in milk samples.

Materials and Methods: A total of 224 subclinical bovine mastitis milk samples were collected from four provinces of Thailand and determined S. aureus using conventional method and also subjected to the screening test, MSA and DNase test. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) among both tests were analyzed and compared to the tube coagulase test (TCT), as reference method. Immunological test by latex agglutination and molecular assay by determined spa gene were also used to identify and differentiate S. aureus.

Results: A total of 130 staphylococci were isolated by selective media, Gram-stain, and catalase test. The number of S. aureus which identified using TCT, MSA and DNase test were 32, 102, and 74 isolates, respectively. All TCT results were correlated to results of latex agglutination and spa gene which were 32 S. aureus. MSA showed 100% sensitivity, 28.57% specificity, 31.37% PPV, and 100% NPV, whereas DNase showed 53.13% sensitivity, 41.84% specificity, 22.97% PPV, and 73.21% NPV. DNase test showed higher specificity value than MSA but the test presented 26.79% false negative results whereas no false-negative result from MSA when comparing to TCT.

Conclusion: MSA had a tendency to be a good preference for screening S. aureus because of its high sensitivity and NPV. The result from this study will improve a choice to use a screening test to diagnose S. aureus of veterinary field for prompt disease controlling and effective treatment.

Keywords: bovine mastitis, deoxyribonuclease test, mannitol fermentation test, screening methods, Staphylococcus aureus.

Virulence gene profiles of Arcobacter species isolated from animals, foods of animal origin, and humans in Andhra Pradesh, India

Research (Published online: 30-06-2017)

23. Virulence gene profiles of Arcobacter species isolated from animals, foods of animal origin, and humans in Andhra Pradesh, India

M. Soma Sekhar, S. R. Tumati, B. K. Chinnam, V. S. Kothapalli and N. Mohammad Sharif

Veterinary World, 10(6): 716-720

ABSTRACT

Aim: This study aimed to detect putative virulence genes in Arcobacter species of animal and human origin.

Materials and Methods: A total of 41 Arcobacter isolates (16 Arcobacter butzleri, 13 Arcobacter cryaerophilus, and 12 Arcobacter skirrowii) isolated from diverse sources such as fecal swabs of livestock (21), raw foods of animal origin (13), and human stool samples (7) were subjected to a set of six uniplex polymerase chain reaction assays targeting Arcobacter putative virulence genes (ciaB, pldA, tlyA, mviN, cadF, and cj1349).

Results: All the six virulence genes were detected among all the 16 A. butzleri isolates. Among the 13 A. cryaerophilus isolates, cadF, ciaB, cj1349, mviN, pldA, and tlyA genes were detected in 61.5, 84.6, 76.9, 76.9, 61.5, and 61.5% of isolates, respectively. Among the 12 A. skirrowii isolates, cadF, ciaB, cj1349, mviN, pldA, and tlyA genes were detected in 50.0, 91.6, 83.3, 66.6, 50, and 50% of isolates, respectively.

Conclusion: Putative virulence genes were detected in majority of the Arcobacter isolates examined. The results signify the potential of Arcobacter species as an emerging foodborne pathogen.

Keywords: Arcobacter, Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, polymerase chain reaction, virulence genes.

Honey can repairing damage of liver tissue due to protein energy malnutrition through induction of endogenous stem cells

Research (Published online: 29-06-2017)

22. Honey can repairing damage of liver tissue due to protein energy malnutrition through induction 

of endogenous stem cells

R. Heru Prasetyo and Eka Pramyrtha Hestianah

Veterinary World, 10(6): 711-715

ABSTRACT

Aim: This study was to evaluate effect of honey in repairing damage of liver tissue due to energy protein malnutrition and in mobilization of endogenous stem cells.

Materials and Methods: Male mice model of degenerative liver was obtained through food fasting but still have drinking water for 5 days. It caused energy protein malnutrition and damage of liver tissue. The administration of 50% (v/v) honey was performed for 10 consecutive days, while the positive control group was fasted and not given honey and the negative control not fasted and without honey. Observations of regeneration the liver tissue based on histologically examination, observation of Hsp70 expression, and homing signal based on vascular endothelial growth factor-1 (VEGF-1) expression using immunohistochemistry technique. Observation on expression of CD34 and CD45 as the marker of auto mobilization of hematopoietic stem cells using flow cytometry technique.

Results: There is regeneration of the liver tissue due to protein energy malnutrition, decrease of Hsp70 expression, increase of VEGF-1 expression, and high expression of CD34 and CD45.

Conclusion: Honey can improve the liver tissue based on: (1) Mobilization of endogenous stem cells (CD34 and CD45); (2) Hsp70 and VEGF-1 expressions as regeneration marker of improvement, and (3) regeneration histologically of liver tissue.

Keywords: endogenous stem cells, honey, liver tissue, protein energy malnutrition, regeneration.

Tetracycline resistance phenotypes and genotypes of coagulase-negative staphylococcal isolates from bubaline mastitis in Egypt

Research (Published online: 25-06-2017)

21. Tetracycline resistance phenotypes and genotypes of coagulase-negative staphylococcal 

isolates from bubaline mastitis in Egypt

K. A. Abd El-Razik, A. A. Arafa, R. H. Hedia and E. S. Ibrahim

Veterinary World, 10(6): 702-710

ABSTRACT

Aim: This study was devoted to elucidate the tetracycline resistance of coagulase-negative staphylococci (CNS) derived from normal and subclinical mastitic (SCM) buffaloes' milk in Egypt.

Materials and Methods: A total of 81 milk samples from 46 normal buffalo milk samples and 35 SCM buffalo milk samples at private dairy farms of Egypt were used in this study. CNS were identified using phenotypic and molecular methods (polymerase chain reaction [PCR]). CNS isolates were tested for tetracycline resistance using routine methods and multiplex PCR targeting tetracycline (tet) resistance genes followed by sequencing of positive PCR products and phylogenetic analysis.

Results: Isolation and identification of 28 (34.5%) CNS from normal and SCM buffaloes' milk, namely, Staphylococcus intermedius (39.2%), Staphylococcus xylosus (25.0%), Staphylococcus epidermidis (10.7%), Staphylococcus hominis (10.7%), and 3.5% to each of Staphylococcus sciuri, Staphylococcus hyicus, Staphylococcus lugdunensis, and Staphylococcus simulans. Using nested PCR, all the 28 CNS isolates revealed positive for 16srRNA gene specific for genus staphylococci and negative for thermonuclease (nuc) gene specific for Staphylococcus aureus species. The presence of tetracycline resistance-encoding genes (tetK, tetL, tetM, and tetO) was detected by multiplex PCR. All isolates were negative for tetL, M, and O genes while 14 (50%) CNS isolates were positive for tetK gene, namely, S. lugdunensis (100%), S. hominis (100%), S. epidermidis (66.6%), S. intermedius(45.4%), and S. xylosus (42.8%). Nucleotide sequencing of tetK gene followed by phylogenetic analysis showed the high homology between our CNS isolates genes of tetracycline resistance with S. aureus isolates including Egyptian ones. This proves the transfer of the tetracycline resistance encoding genes between coagulase-negative and coagulase-positive Staphylococcus spp.

Conclusion: CNS isolates have distinguishingly high resistance to tetracycline. Abundant tetracycline usage for mastitis treatment leads to the spread of genetic resistance mechanisms inside CNS strains and among all Staphylococcus spp. Consequently, tetracycline is not effective anymore.

Keywords: buffaloes, coagulase-negative staphylococci, mastitis, tetracycline resistance, tetK gene.