Evaluation of chimeric proteins for serological diagnosis of brucellosis in cattle

Research (Published online: 24-08-2021)

27. Evaluation of chimeric proteins for serological diagnosis of brucellosis in cattle

Aitbay K. Bulashev, Bakytkali K. Ingirbay, Kanatbek N. Mukantayev and Alfiya S. Syzdykova

Veterinary World, 14(8): 2187-2196

ABSTRACT

Background and Aim: An accurate diagnosis of Brucella-infected animals is one of the critical measures in eradication programs. Conventional serological tests based on whole-cell (WC) antigens and detecting antibodies against pathogen-associated lipopolysaccharide might give false-positive results due to the cross-reactivity with other closely related bacteria. This study evaluated the serological potential of Brucella spp. chimeric outer membrane proteins (Omps) as antigens in an indirect enzyme-linked immunosorbent assay (i-ELISA).

Materials and Methods: The chimeric gene constructs of the most immunodominant regions of Brucella Omps 25+31, 25+19, and 19+31 were cloned into the pET28a expression vectors and transformed into Escherichia coli BL21 (DE3). The serological potential of chimeric proteins compared with single recombinant Omps (rOmps)19, 25, and/or 31 were studied on blood serum samples of (i) a rabbit immunized with killed Brucella abortus 19WC, (ii) mice immunized with single rOmps, (iii) cows seropositive for brucellosis by rose Bengal test, and (iv) cattle naturally and/or experimentally infected with brucellosis.

Results: E. coli BL21 actively produced Brucella chimeric rOmps, the concentration of which reached a maximum level at 6 h after isopropyl-β-D-1-thiogalactopyranoside stimulation. Target proteins were antigenic and expressed in an active state, as recognized by rabbit anti-B. abortus antibodies in an i-ELISA and western blotting. Murine antibodies against the single rOmps reacted with chimeric antigens, and conversely, antichimeric antibodies found their epitopes in single proteins. Brucella chimeric rOmps showed higher antigenicity in blood sera of seropositive cattle kept in the hotbed of the infection and/or experimentally challenged with brucellosis than single proteins.

Conclusion: Brucella chimeric recombinant outer membrane proteins could be a potential antigen candidate for developing an ELISA test for accurate diagnosis of bovine brucellosis.

Keywords: Brucella, chimeric protein, enzyme-linked immunosorbent assay, serological potential.

Knowledge, attitude, and practices associated with rabies in villages with different dog vaccination statuses in Cambodia

Research (Published online: 24-08-2021)

26. Knowledge, attitude, and practices associated with rabies in villages with different dog vaccination statuses in Cambodia

Bunsong Ung, Ketsarin Kamyingkird and Waraphon Phimpraphai

Veterinary World, 14(8): 2178-2186

ABSTRACT

Background and Aim: Rabies is a highly infectious but neglected zoonotic disease. Almost 99% of rabies-related human deaths are caused by dog-mediated rabies. Although canine rabies vaccination is highly effective and provides protection, nationwide rabies vaccination campaigns have been insufficient in Cambodia, resulting in a limited number of rabies vaccinated dogs. This study aimed to explore the rabies knowledge, attitude, and practices (KAP) among participants from both dog rabies vaccinated (DRV) and dog rabies unvaccinated (DRUV) villages located in the Kandal and Prey Veng Provinces, Cambodia.

Materials and Methods: A cross-sectional survey was conducted with dog owners in Kandal and Prey Veng Provinces, Cambodia, during August and September 2020. The structural questionnaire collected general sociodemographic information and the KAP associated with rabies transmission, clinical signs, management, and control. The data were then analyzed using Wilcoxon rank-sum test and Chi-square statistics.

Results: In total, 312 participants were interviewed: 137 participants from DRV villages and 175 from DRUV villages. Among them, 99.4% (310/312) had previously heard about rabies. Out of these 310, 93.5% (290/310) were aware that rabies is a fatal disease, while 96.5% (299/310) were willing to vaccinate their dog against rabies if the vaccination was provided for free. However, 32.9% (102/310) indicated that they would be willing to sell their own dog if it bit someone or showed aggression. More than one-third (115/310) of all the respondents had poor overall KAP regarding rabies. The respondents from DRV villages had significantly higher overall scores with regard to rabies KAP than those from DRUV villages (p<0.0001). According to the factors related to overall KAP, village type and education level were significantly associated with overall KAP of the respondents (p<0.0001).

Conclusion: The rabies disease is recognized in Cambodia, and dog owners are willing to vaccinate their dogs if the vaccination is provided for free. The overall rabies-related KAP were poor among 30% of the respondents, and higher KAP scores were obtained for the DRV villages. The village type and education level were found to be associated with the different overall KAP of the participants.

Keywords: attitude, knowledge, practices, rabies.

Comparative study of the therapeutic efficacy of autologous platelet-rich plasma and honey in healing skin wounds in sheep

Research (Published online: 23-08-2021)

25. Comparative study of the therapeutic efficacy of autologous platelet-rich plasma and honey in healing skin wounds in sheep

Daikh Badis and Deffa Ouafa

Veterinary World, 14(8): 2170-2177

ABSTRACT

Background and Aim: This investigation is the continuation of a published preliminary study examining the therapeutic efficacy of platelet-rich plasma (PRP) as a topical treatment for skin wounds in sheep. The study aimed to compare the healing effects of autologous PRP with that of natural honey.

Materials and Methods: This study involved nine clinically healthy male sheep. After sterile skin preparation, full-thickness longitudinal incision wounds were created on the backs of each animal. The animals were randomly divided into three groups of three sheep each. In Group I, the wounds were treated with PRP; in Group II, the wounds were treated with honey; and in Group III, the wounds were treated with saline solution. The different treatments were administered topically every 3 days. Healing was assessed by a semi-quantitative histopathological study from biopsies taken on the 3rd, 7th, 14th, 21st, and 28th days of healing. The data obtained were compared using the non-parametric Mann–Whitney U-test, and p<0.05 and 0.01 were used to determine the level of significance of the recorded differences.

Results: Semi-quantitative histopathological evaluation showed significant differences in the progression of wound healing between the three study groups. Recorded data showed that PRP may reduce inflammation during the first 3 days after the incision. Moreover, the synthesis and organization of collagen fibers were significantly improved in the group treated with PRP compared with those in the group treated with honey.

Conclusion: PRP offers a promising therapeutic option for healing skin wounds in sheep compared with honey.

Keywords: honey, platelet-rich plasma, sheep, skin, wound healing.

A parasitological survey of zoonotic cestodes carried by house rats in Aswan, Egypt, reveals cryptic diversity at the molecular level

Research (Published online: 23-08-2021)

24. A parasitological survey of zoonotic cestodes carried by house rats in Aswan, Egypt, reveals cryptic diversity at the molecular level

Abuelhassan Elshazly Younis, Atef Ibrahim Saad, Islam Refaat Mohamed El-Akhal and Nagla Mustafa Kamel Saleh

Veterinary World, 14(8): 2160-2169

ABSTRACT

Background and Aim: Some rat cestodes are zoonotic and are capable of parasitizing humans and animals, raising serious concerns regarding human and veterinary health. The study aimed to determine the prevalence and risk factors for cestodes in Egyptian house rats and to characterize the cestodes molecularly.

Materials and Methods: The current survey examined 115 house rats (Rattus rattus) in two cities (Edfu and Aswan) in Egypt's Aswan Governorate for cestode infection using integrated molecular approaches (polymerase chain reaction, sequencing, and phylogenetic analysis) and morphological/morphometrical approaches.

Results: The cestodes identified in this study exhibited the typical morphological characteristics of Hymenolepis diminuta (Rudolphi, 1819), Hymenolepis nana (Siebold, 1852) (from rat intestine), and Hydatigera taeniaeformis (from rat liver). The species prevalence rates from these three studies were reported to be 8.7%, 10.4%, and 20.9%, respectively. The ribosomal DNA (ITS1, 18S, and complete ITS) sequences revealed that the hymenolepid sequences were highly distinct but were related to other sequences in the GenBank database, with some sequences showing high similarities to those of H. nana and H. diminuta. In addition, the H. taeniaeformis sequences (ITS2 and mitochondrial cytochrome c oxidase subunit 1 [mtCOX1]) obtained in this study were highly similar to some Taenia taeniaeformis GenBank sequences. The constructed phylogram revealed that the hymenolepidid tapeworms examined in this study were classified into four major branches (the majority of which were hybrids of the two species) and belonged to the genus Hymenolepis. In addition, the phylogram of H. taeniaeformis assigned this species to T. taeniaeformis.

Conclusion: When typical hymenolepid morphology is combined with molecular and phylogenetic divergence, it may indicate the existence of possible cryptic species. In addition, on the basis of the phylogenetic analysis, genetic diversity within T. taeniaeformis may exist as determined by comparing the metacestode mtCOX1 sequences. The current study presents the prevalence values of zoonotic cestodes and contributes to the body of knowledge, including identification keys and the use of molecular tools for species confirmation.

Keywords: cestodes, cryptic diversity, molecular identification, rat.

New phenomena for clinicians, model of Candida albicans mobilization before and after biofilm formation in the intestinal mucosa of Wistar rats (Rattus norvegicus)

Research (Published online: 23-08-2021)

3. New phenomena for clinicians, model of Candida albicans mobilization before and after biofilm formation in the intestinal mucosa of Wistar rats (Rattus norvegicus)

Masfufatun Masfufatun, Loo Hariyanto Raharjo, Harsono Wiradinata, Putu Oky Ari Tania, Ni'matuzahroh Ni'matuzahroh and Afaf Baktir

International Journal of One Health, 7(2): 165-170

ABSTRACT

Background and Aim: The virulence and antifungal resistance of Candida albicans are recently known for their ability to form biofilm. This research aimed to construct an in vivo model of C. albicans biofilm in Wistar rats' intestinal mucosa and study their mobilization while in a planktonic and biofilm formation. In this study, there was one treatment group that was treated with three antibiotics, immunosuppressants, and C. albicans.

Materials and Methods: This study was divided into control and treatment groups. The data sampling was conducted after C. albicans inoculation. The C. albicans biofilm formation stage was monitored with colony-forming units method calculation every week post-inoculation and then observed by the confocal laser scanning microscope.

Results: The planktonic C. albicans overgrowth occurred up to 14 days after inoculation. The formation and maturation of C. albicans biofilm in the intestinal mucosa started in the 28th and 35th-day post-inoculation, respectively. The density of planktonic C. albicans in the stool was dramatically decreased on the 35th day. Before the biofilm formation, the planktonic Candida was carried away by food scraps to be released as a stool. However, there were minuscule or no planktonic Candida observed in the stool during and after biofilm formation. Instead, they were attached to the caecum's mucosa as a biofilm.

Conclusion: We have proved that the planktonic C. albicans with its mobile nature were carried into the stool along with the rest of the feed, as we observed a lot of C. albicans cells found in the stool. Meanwhile, on day 28 after administration of antibiotics and immunosuppressants, no C. albicans was found in the stool samples, and at the same time, we observed C. albicans cells and their matrix attached to the intestinal mucosa as a biofilm.

Keywords: biofilm, candidiasis, Candida albicans, confocal laser scanning microscope, intestinal mucosa.

Pseudomonas aeruginosa infection of avian origin: Zoonosis and one health implications

Review (Published online: 21-08-2021)

23. Pseudomonas aeruginosa infection of avian origin: Zoonosis and one health implications

Wafaa A. Abd El-Ghany

Veterinary World, 14(8): 2155-2159

ABSTRACT

Zoonotic diseases are diseases that are transmitted from animals to humans and vice versa. Pseudomonas aeruginosa (P. aeruginosa) is a pathogen with zoonotic nature. Commercial poultry could be infected with P. aeruginosa, especially at young ages with great losses. Infection of embryos with P. aeruginosa induced death in the shell, while infection of chicks led to septicemia, respiratory and enteric infections, and high mortality. Humans are also highly susceptible to P. aeruginosa infection, and the disease is associated with severe lung damage, especially in immunocompromised patients. Chicken carcass and related poultry retail products play an important role in the transmission of P. aeruginosa to humans, especially after processing in abattoirs. Treatment of P. aeruginosa infection is extremely difficult due to continuous development of antibiotic resistance. The transfer of antibiotic-resistant genes from poultry products to humans creates an additional public health problem. Accordingly, this study focused on avian pseudomonad, especially P. aeruginosa, with respect to infection of poultry, transmission to humans, and treatment and antibiotic resistance.

Keywords: antibiotic resistance, human, poultry, Pseudomonas aeruginosa, zoonosis.

Characterization of serum protein electrophoresis patterns and C-reactive protein in canine tick-borne diseases

Research (Published online: 21-08-2021)

22. Characterization of serum protein electrophoresis patterns and C-reactive protein in canine tick-borne diseases

Sariya Asawakarn and Piyanan Taweethavonsawat

Veterinary World, 14(8): 2150-2154

ABSTRACT

Background and Aim: Canine tick-borne diseases are important diseases with a worldwide distribution. In Thailand, the most important canine tick-borne diseases are ehrlichiosis, babesiosis, and hepatozoonosis. This study aimed to determine the serum protein electrophoresis patterns (SPEPs) and C-reactive protein (CRP) levels associated with Ehrlichia canis, Babesia canis, or Hepatozoon canis single infections.

Materials and Methods: A total of 650 canine blood samples were collected from animal hospitals and clinics in Bangkok and its vicinity to examine health status and blood parasite infection. Suspected blood parasite infections were examined by buffy coat thin blood smear and confirmed by polymerase chain reaction. Normal dog and positive E. canis, B. canis, and H. canis single infections and serum protein profiles were determined by agarose gel electrophoresis. CRP concentration was measured by fluorescent immunoassay.

Results: In dogs infected with E. canis, B. canis, and H. canis single infections, albumin levels and A/G ratios significantly decreased, whereas β2-globulin levels increased (p<0.05). The γ-globulin level significantly increased in E. canis and H. canis infections (p<0.05). A monoclonal gammopathy pattern was observed in E. canis and B. canis single infections, whereas β-γ bridging patterns and increased β- and γ-globulin fractions were found in H. canis single infections. The CRP level increased in dogs with blood parasite single infections and may be related to the pathogenesis of the infection.

Conclusion: SPEPs and CRP levels can be used to monitor health status and blood parasite problems in infected dogs.

Keywords: blood parasites, C-reactive protein, dog, electrophoresis, serum protein.