Vibriosis and Aeromonas infection in shrimp: Isolation, sequencing, and control

Research (Published online: 14-05-2019)

6. Vibriosis and Aeromonas infection in shrimp: Isolation, sequencing, and control

Hanaa Mohamed Fadel and Maather El-Lamie M.M.

International Journal of One Health, 5: 38-48

Background and Aim: Shrimp is one of the most commonly consumed types of seafood. It is a very nutritious healthy food. Shrimp is low in calories and rich in protein and healthy fats. It also contains a treasure trove of vitamins and minerals. On the negative side, it may be affected by many bacterial diseases which affect its health. Furthermore, it may be incriminated as a vector of foodborne illnesses that range from mild gastrointestinal upset to life-threatening diseases. This study was designed to assess the clinical picture and zoonotic importance of vibriosis and Aeromonas infection in live shrimp and to study the antibacterial effect of citric acid (lemon juice) and acetic acid (vinegar) on these pathogens.

Materials and Methods: A total of 170 live shrimp (Metapenaeus monoceros) samples were collected from Suez City, Egypt. The samples were examined clinically, and then, they were enriched into alkaline peptone water and cultivated on thiosulfate-citrate-bile salts-sucrose agar and ampicillin MacConkey agar for the isolation of Vibrio and Aeromonas species, respectively. The recovered isolates were confirmed biochemically and genotypically using duplex polymerase chain reaction (PCR) and sequencing. The germicidal effects of vinegar and lemon on artificially contaminated shrimp samples with Aeromonas hydrophila and Vibrio parahaemolyticus at different times (0.25, 1, 1.5, and 24 h) and temperatures (5° and 30°C) were studied.

Results: The results revealed that some of the infected shrimp were hypoxic, lethargic with abnormal swimming behavior. In most cases, body appendages, telsons, uropods, and gills took black coloration. In addition, the hepatopancreas appeared soft, swollen, and congested. The prevalence rates of vibriosis in each of the musculature and hepatopancreas were 4.7%, while the prevalence rates of Aeromonas infection in the musculature and hepatopancreas were 11.8% and 11.2%, respectively. Duplex PCR showed that Aeromonas isolates gave double bands: 237 bp specific for gcat and 500 bp specific for 16S rRNA, while Vibrio spp. and Plesiomonas shigelloides isolates gave single band at 500 bp. The effect of organic acid treatment showed that acetic acid (vinegar 5%) had increasing reduction rates that reached its maximum level after 24 h; where it caused (100% inhibition) for A. hydrophila at both temperatures and (33.63% and 60% inhibition) for V. parahaemolyticus at refrigerator and room temperatures, respectively. Moreover, acetic acid was more effective at room temperature than at refrigerator temperature. Concerning the effect of lemon juice (citric acid), it was more effective than acetic acid at short marination (0.25 and 1 h) at both temperatures for the two pathogens. Moreover, lemon was more effective at refrigerator temperature than at room temperature at the same aforementioned time. The difference between the reduction effects of the two acids on both pathogens was statistically significant (p<0.0001).

Conclusion: Overall, the examined shrimp samples were found to be vectors for Vibrio and Aeromonas spp. Application of hygienic measures during handling and cooking of shrimp should be esteemed. The organic acid treatment trial showed that vinegar and lemon juice can be used as a safe and economic method to limit the microbial contamination in seafood.

Keywords: Shrimp, vibriosis, Aeromonas infection, sequencing, decontamination, citric acid, acetic acid.

Isolation and molecular characterization of Mycoplasma spp. in sheep and goats in Egypt

Research (Published online: 13-05-2019)

6. Isolation and molecular characterization of Mycoplasma spp. in sheep and goats in Egypt

Mounier M. Abdel Halium, Fayez A. Salib, S. A. Marouf and Emil S. Abdel Massieh

Veterinary World, 12(5): 664-670

ABSTRACT

Background and Aim: Different species of Mycoplasma are associated with many pathological problems in small ruminants including respiratory manifestation, this problem results in significant losses, especially in African countries. This study aimed to (I) study some epidemiological aspects of Mycoplasma species infections in Egyptian sheep and goats at Giza Governorate, (II) diagnosis of Mycoplasma species affections using bacterial isolation and identification, (III) apply the polymerase chain reaction (PCR) for typing of different Mycoplasma species, and (IV) illustrate the phylogenetic tree for the isolated Mycoplasma species and other species from GenBank using the purified PCR product.

Materials and Methods: A total of 335 samples were collected from sheep and goats from Giza Governorate in Egypt as 142 nasal swabs from clinically affected animals, 167 pneumonic lungs, 18 samples from tracheal bifurcation, and 8 samples by bronchial wash were cultured on pleuropneumonia-like organisms (PPLOs) media for cultivation of Mycoplasma species. PCR and sequencing and phylogenetic analysis were adopted to identify and classify the isolated Mycoplasma species.

Results: A total of 24 Mycoplasma isolates were isolated on PPLO media, identified by biochemical tests, and confirmed and typed by PCR using specific primers. 10 isolates were confirmed as Mycoplasma arginini, four isolates as Mycoplasma ovipneumoniae by PCR, and 10 isolates as undifferentiated Mycoplasma species. A purified isolate of M. arginini and M. ovipneumoniae was sequenced and phylogenetic analysis was illustrated.

Conclusion: M. arginini and M. ovipneumoniae are prevalent in Egyptian sheep and goats. Further studies on M. arginini are required due to its high frequency of isolation from pneumonic sheep and goats and also from animals suffer from different respiratory manifestations.

Keywords: goats, Mycoplasma, polymerase chain reaction, sequencing and phylogenetic analysis, sheep.

Medicinal plants and their effects on diabetic wound healing

Review (Published online: 11-05-2019)

5. Medicinal plants and their effects on diabetic wound healing

Oluwafemi O. Oguntibeju

Veterinary World, 12(5): 653-663

ABSTRACT

Wounds have a serious negative impact on the health-care economy of a country, especially on the economy of developing countries where resources are poor and funding is very limited. It is presumed that about 80% of people living in developing countries use traditional medicines which are majorly prepared from medicinal plants to meet their primary health-care needs. Due to the large reservoir of medicinal plants and adequate traditional knowledge on wound healing, many people in Africa and other developing countries use medicinal plants in the treatment of diabetic wounds and related complications. Wound healing in the external and internal biological organs involves a series of complex overlapping processes which demand excellent communication between cells. It is an orderly and highly controlled process characterized by hemostasis, inflammation, proliferation, and remodeling. Diabetes is a global health problem predicted to rise to over 642 million by 2040. The propelling factor responsible for the increase in morbidity and mortality of diabetes is linked to vascular complications as well as the failure of the wound healing processes in diabetic state. Different approaches have been adopted in the treatment of diabetic wounds, and medicinal plants are certainly one of those approaches that have drawn global attention. In this review paper, the effects of medical plants on wound healing in diabetic state as well as factors affecting wound healing and the mechanism of action of medicinal plants are examined.

Keywords: Diabetes, healing, infection, medicinal plants, wounds.

The prevalence of horse trypanosomiasis in Sumba Island, Indonesia and its detection using card agglutination tests

Research (Published online: 09-05-2019)

4. The prevalence of horse trypanosomiasis in Sumba Island, Indonesia and its detection using card agglutination tests

Wisnu Nurcahyo, Marlin R. K. Yowi, Sri Hartati and Joko Prastowo

Veterinary World, 12(5): 646-652

ABSTRACT

Background and Aim: Horses have a strategic and vital role to play in the lives of the people of Sumba Island, East Nusa Tenggara Province. They act as social animals that are involved in death ceremonies, horse races, and during pasola, thereby supporting tourism, and are given away as dowry in wedding ceremonies. This study aimed to investigate the prevalence of trypanosomiasis among horses in four districts of Sumba Island by examining clinical symptoms and detecting parasites, antibodies, and other factors that are related to Trypanosoma evansi infection in horses.

Materials and Methods: We studied a total of 211 horses that belonged to 88 clinical hobby breeders. Giemsa-colored smears and serum were examined in order to detect antibodies using card-agglutination tests (CATT). The study was conducted during the rainy season that lasted from January to March 2017. Potential risk factors such as the species, sex, origin of the livestock, how the livestock were maintained, and the farmers' knowledge concerning trypanosomiasis were recorded using questionnaires. Data were collected annually for three years from 2010-2012 and repeatedly analyzed by a Chi-square test.

Results: Clinical signs of trypanosomiasis were found in 34 horses; blood smears were examined using Giemsa staining and negative preparations were obtained at a frequency of 0.0% (0/211). The CATT results generally showed that 13.3% (28/211) of the samples were seropositive for antibodies to T. evansi; the highest percentage, 16.67% (8/48), of seropositivity was found in the West Sumba District, and the lowest, 12.0% (5/50), was found in Southwest Sumba. The incidence of trypanosomiasis was higher (75% [21/28]) among female hip horses; horses with 1-5 years of experience were more susceptible to a T. evansi infection (46.4% [13/28]). In general, farmers on Sumba Island knew of trypanosomiasis (89.8% [79/88]), and 69.3% (61/88) of the farmers reported that their livestock was sick. This study was the first serological study conducted on trypanosomiasis in horses of Sumba Island after the surra outbreak in 2010-2012. There were 3% of farmers who were willing to provide the government with information on implementing a prevention program and controlling the spread of surra on the island.

Conclusion: The diagnoses of surra disease were made based on clinical symptoms and parasitological examinations. CATTs could be used to diagnose T. evansi infection in horses.

Keywords: CATT, horses, Sumba Island, Trypanosoma evansi.

Study on prevalence, clinical presentation, and associated bacterial pathogens of goat mastitis in Bauchi, Plateau, and Edo states, Nigeria

Research (Published online: 07-05-2019)

3. Study on prevalence, clinical presentation, and associated bacterial pathogens of goat mastitis in Bauchi, Plateau, and Edo states, Nigeria

Faruq Ahmad Danmallam and Nikolai Vasilyevich Pimenov

Veterinary World, 12(5): 638-645

ABSTRACT

Aim: This study aimed to estimate the prevalence and clinical presentations of different forms of mastitis and mastitis-causing pathogens in lactating goats in Bauchi, Plateau, and Edo states, Nigeria.

Materials and Methods: A total of 500 quarters from 250 lactating goats of Red Sokoto and West African Dwarf breeds during the lactation period were clinically examined. Clinical mastitis was detected by gross signs of udder infection during physical examination and abnormal milk, whereas subclinical mastitis (SCM) was recognized using California mastitis test. The bacterial pathogens were identified by morphology, hemolysis, gram staining, and biochemical tests such as catalase, oxidase, coagulase, reaction on sulfite, indole, and motile medium, and fermentation of sugars.

Results: The overall prevalence of mastitis in goats was found to be 101 (40.4%), of which 8% (20/250) were clinical, and 32.4% (81/250) were SCM cases. The quarter level prevalence was 29.4% (145/493), comprising 5.9% (29/493) clinical and 23.2% (116/493) subclinical forms of mastitis. In addition, 1.4% (7/500) of teats were found to be blind on the clinical examination of the udder and teat. Several regional inflammatory reactions and abnormalities in milk were found in 69% and 100% of the cases, respectively. Moreover, some indications of generalized signs such as fever, reduction in appetite, increase in respiration, and pulse rate per minute were recorded in 100%, 75%, 85% and 80% of the cases, respectively. The predominant bacterial isolates recovered were Staphylococcus aureus (20.0%), followed by Escherichia coli (15.5%) and Streptococcus agalactiae (11.0%), and the least isolated microorganisms (≤6%) were bacteria of different species including Staphylococcus auricularis, Staphylococcus caprae, Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus hyicus, Staphylococcus xylosus, Staphylococcus lentus, Streptococcus dysgalactiae, Streptococcus pluranimalium, Streptococcus uberis, Streptococcus pneumoniae, Streptococcus ruminatorum, Streptococcus suis, Micrococcus luteus, Enterobacter cloacae, Proteus vulgaris, Klebsiella oxytoca, Klebsiella pneumoniae, Morganella morganii, Salmonella Typhimurium, Citrobacter freundii, Pseudomonas aeruginosa, Acinetobacter rudis, Acinetobacter haemolyticus, and Bacillus cereus.

Conclusion: Mastitis continues to be recognized as one of the important health issues and leads to major economic losses to the dairy goats caused by many bacterial pathogens, and the effective measures need to be taken to control the disease.

Keywords: bacteria, goats, mastitis, Nigeria, physical feature, prevalence.

Screening commercial teat disinfectants against bacteria isolated from bovine milk using disk diffusion

Research (Published online: 06-05-2019)

2. Screening commercial teat disinfectants against bacteria isolated from bovine milk using disk diffusion

Sarah Rose Fitzpatrick, Mary Garvey, Kieran Jordan, Jim Flynn, Bernadette O'Brien and David Gleeson

Veterinary World, 12(5): 629-637

ABSTRACT

Background and Aim: Teat disinfection is an important tool in reducing the incidence of bovine mastitis. Identifying the potential mastitis-causing bacterial species in milk can be the first step in choosing the correct teat disinfectant product. The objective of this study was to screen commercial teat disinfectants for inhibition against mastitis-associated bacteria isolated from various types of milk samples.

Materials and Methods: Twelve commercially available teat disinfectant products were tested, against 12 mastitis-associated bacteria strains isolated from bulk tank milk samples and bacterial strains isolated from clinical (n=2) and subclinical (n=3) quarter foremilk samples using the disk diffusion method.

Results: There was a significant variation (7-30 mm) in bacterial inhibition between teat disinfection products, with products containing a lactic acid combination (with chlorhexidine or salicylic acid) resulting in the greatest levels of bacterial inhibition against all tested bacteria (p<0.05).

Conclusion: In this study, combined ingredients in teat disinfection products had greater levels of bacterial inhibition than when the ingredients were used individually. The disk diffusion assay is a suitable screening method to effectively differentiate the bacterial inhibition of different teat disinfectant products.

Keywords: bacteria, disk diffusion, mastitis, teat disinfectant.

Electrophoretic profile of seminal proteins and their correlation with in vitro sperm characters in Black Bengal buck semen

Research (Published online: 03-05-2019)

1. Electrophoretic profile of seminal proteins and their correlation with in vitro sperm characters in Black Bengal buck semen

M. Karunakaran, Vivek C. Gajare, Ajoy Mandal, Mohan Mondal, S. K. Das, M. K. Ghosh, S. Rai and R. Behera

Veterinary World, 12(5): 621-628

ABSTRACT

Aim: This study aimed to study the electrophoretic properties of seminal plasma and sperm proteins of Black Bengal buck semen and their correlation with in vitro sperm characters and freezability.

Materials and Methods: Semen ejaculates from nine Black Bengal bucks were collected by artificial vagina (n=20/buck). Ejaculates were evaluated for in vitro sperm characters and electrophoretic profile of seminal protein. In vitro sperm characters were evaluated immediately after collection, after completion of equilibration period, and after freeze-thawing. For seminal protein studies, seminal plasma proteins were precipitated by ice-cold ethanol method, and sperm proteins were extracted by Triton X detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to assess the molecular weight of seminal proteins. Correlation between in vitro sperm characters and protein bands was determined by Pearson's correlation coefficient, and two-way ANOVA was applied to find the individual buck differences.

Results: Significant difference (p<0.01) among the bucks was noticed in the in vitro sperm characters evaluated at all the three stages of semen evaluation such as immediately after collection, after completion of equilibration period, and post-freeze thawing. Progressive loss of sperm motility, membrane integrity, and other in vitro sperm characters were noticed during cryopreservation. A total of ten protein bands in the molecular weight ranging from 17 to 180 kDa were found in the SDS-PAGE of seminal plasma proteins, while nine bands of 17-134 kDa were observed in sperm proteins. Seminal plasma proteins of molecular weight 75, 62-49, 20, and 17 kDa and sperm proteins of 75, 20, and 17 kDa were present in all the nine bucks (100%) screened, and variation among the bucks was noticed for the presence of other proteins. Seminal plasma protein of 180-134 kDa showed a negative correlation with individual motility (−0.716) and functional membrane integrity of sperm cells (−0.724) in post-freeze-thaw analysis and 48 kDa protein had a positive correlation with individual motility (0.649) and functional membrane integrity of sperm cells (0.664) in post-thaw analysis. Sperm proteins of 63 kDa had a negative correlation (−0.616) with sperm concentration in neat semen.

Conclusion: Variation among the bucks was noticed in the in vitro sperm characters and semen freezability. Correlation between seminal proteins and in vitro sperm characters and semen freezability had been found which might be useful as a tool to select breeding bucks.

Keywords: buck, in vitro characters, semen, seminal proteins.