Serological survey of Toxoplasma gondii infection in the one-humped camels (Camelus dromedarius) in the south of Iran.

BACKGROUND: Toxoplasmosis, an important zoonotic disease, is caused by Toxoplasma gondii. Camels are one of several host species for T. gondii parasites and play an important role in the transmission of T. gondii to humans. OBJECTIVES: The present study aimed to describe the seroprevalence of T. gondii in dromedary camels (Camelus dromedarius) from three provinces (Fars, Bushehr and Hormozgan), southern Iran first for this host. METHODS: A total of 180 serum samples were analysed for the presence of anti-Toxoplasma IgG antibodies using the enzyme-linked immune-sorbent assay. RESULTS: Our results showed an overall seroprevalence of T. gondii in 15% of animals. Antibodies to T. gondii were found in sera of 27 of 180 dromedary camels from Fars, Bushehr and Hormozgan provinces, southern Iran. Age or the gender of the camel did not significantly affect the seroprevalence (p > 0.05). There was no significant association between herd-level seroprevalence of T. gondii infection and abortion history, province location residence, history of animal keeping and history of contact with other animals (p > 0.05). CONCLUSIONS: The results of this study showed the presence of T. gondii antibodies among camels in Southern Iran, which could be a public health concern. According to the prevalence of T. gondii infection in camel, the implementation of control measures to reduce infection in both definitive and intermediate hosts is needed.

Molecular characterization and phylogenetic analysis of Linguatula serrata isolated from camels, sheep and goats in Iran.

Linguatula serrata is an important zoonotic parasite with worldwide distribution. The objective of the present study was to investigate the molecular characterization and phylogenetic analysis of nymphal stage of L. serrata from camels, goats and sheep in Iran. The mesenteric lymph nodes were collected from various ruminants including goats, sheep and camels at Isfahan and Shiraz slaughterhouses and the nymphs were identified using morphological characteristics. After DNA extraction, the 18 S rRNA and Cox1 genes were amplified by polymerase chain reaction. The sequencing of the genes was conducted using specific primers and a capillary DNA analyzer. The comparison of amplified sequences with existing data confirmed the presence of L. serrata with 99.6-100% nucleotide sequence similarity. Based on 18 S rRNA and Cox1 sequences, two isolates collected from sheep revealed 100% and 99.9% sequence identity, respectively. Also, three isolates from camel had 99.64-100% and 99.7-100% homology. Two isolates from sheep had 100% identity in their 18SrRNA gene and were categorized together, but showed 99.9% similarity in the Cox1 gene, not clustering together. Phylogenetic analysis of the Cox1 gene classified nearly all the isolates into L. arctica clade. It can be concluded that 18 S rRNA and Cox1 genes sequencing can be a proper method for the analysis of phylogenetic relationships of L. serrata among different hosts in different parts of Iran, possibly helpful for infection control and prevention.

Indigenous production, characterization and evaluation of polyclonal antibody against Camelus dromedarius immunoglobulin.

No diagnostic kits and reagents are available in the market to detect and evaluate camel immune responses to different pathogens. This study aimed to produce sheep anti-camel (Camelus dromedarius) polyclonal antibodies (pAbs) and to determine the specificity with other species immunoglobulin. Immunoglobulins (Igs) from camel serum samples were purified using ammonium sulfate precipitation (40.00% saturated ammonium sulfate). Purity of the camel Igs was tested by sodium dodecyl sulfate polyacrylamide gel electrophoresis. PAbs against (Camelus dromedarius) immunoglobulins were generated by immunizing sheep with purified Igs. Anti- camel Ig polyclonal antibodies titer and specificity were determined using ELISA and Western blot techniques. Polyclonal antibodies specific to camel Igs were significantly high in immunized sheep which confirmed the immunization procedure. PAbs reacted specifically with camel serum immunoglobulin and did not react with other species immunoglobulin of horse and chickens. Polyclonal antibodies produced in this study can be regarded as a valuable tool to be used for immune-diagnostic purposes in camel population world-wide.

Development and evaluation of an indirect ELISA for detection of Teladorsagia circumcincta infection in sheep.

BACKGROUND: The gastrointestinal helminth, Teladorsagia circumcincta, is one of the major health risks and production-limiting diseases in small ruminant populations, particularly in temperate regions. With the increasing importance of disease management and recruited anthelmintic resistant types, accurate approaches are needed for the diagnosis of the infection in the host. Due to uncertain results using faecal examinations, the ELISA method was indicated for the detection of nematode antigenic materials. Despite some promising results, problems were described in terms of test specificity and cross-reactions. Therefore, this study aimed to evaluate the IgG response to worm somatic and excretory/secretory (ES) products using western blot analysis and an indirect ELISA for the detection of T. circumcincta infection in sheep. RESULTS: Based on the immuno-reactivity analysis, immunogenic fractions with molecular weights (MWs) of approximately 60, 75 and 100 kDa were detected in somatic content and two antigens of about 63 and 75 kDa in ES material. Accordingly, a specific product at 75 kDa had the strongest reaction and appeared as the most common antigenic protein. In ELISA, all the sera from the infected sheep revealed the OD rates above the calculated cut-off value with about two-fold greater average. Negative control samples were also specifically recognized with the mean OD rate of about 1/3 of the estimated cut-off value. The cross-reaction test, using rabbit anti-T. circumcincta IgG, did not show reactivity with the ES antigens of other prevalent nematodes including Haemonchus contortus, Protostrongylus rufescens and Marshallagia marshalli. In contrast, a strong positive reaction was observed with the somatic antigens of M. marshalli. CONCLUSIONS: The results of this study indicated that the indirect ELISA method using the ES content enables distinguishing the T. circumcincta infected sheep with high specificity. Those antigenic ES peptides with 63 and particularly 75 kDa MWs should be further investigated due to the potential for serological diagnostic methods and immunoprotective targets in the host.

Effects of two probiotic spores of Bacillus species on hematological, biochemical, and inflammatory parameters in Salmonella Typhimurium infected rats.

Salmonella infections have become a major health concern in recent decades. This pathogen has evolved to become resistant to antibiotics, which has caused problems in its treatment. As such, finding a novel preventive method is important in the treatment and management of this infection. In recent years, uses of probiotics, especially spore-former genera such as Bacillus spp. has become increasingly popular. In this study spores of two probiotic bacteria, Bacillus subtilis and Bacillus coagulans were fed to rats for three weeks through their daily water intake after which Salmonella Typhimurium was gavaged to the rats. On days 1, 3, 5 and 7 after gavaging, the number of Salmonella was counted in liver, spleen, mesenteric lymph nodes, feces and content of ileum and cecum. Hematological and biochemical parameters, inflammatory mediators, total antioxidant capacity and malondialdehyde were also measured. The results showed that B. subtilis and B. coagulans caused delation in infiltration of Salmonella into the lymph nodes, spleen and liver, reduction of the inflammatory mediators, and decreases in oxidative stress, hematological and biochemical changes. The overall count of Salmonella in the above mentioned parameters has also decreased and a faster return to normal base were also witnessed. The results showed that the use of B. subtilis and B. coagulans can potentially help boost the body's immune system, to combat the effects of exposure to the Salmonella pathogen.

Molecular characterization of ß-tubulin gene associated with benzimidazole resistance in larvae of field isolates of Parascaris (Nematoda: Ascarididae).

Since the past 2 decades, an increasing number of resistance to the Benzimidazoles (BZs) have been reported in nematode parasites of livestock. More recently, detection of single-nucleotide polymorphisms (SNPs) at codons of 167, 198 or 200 of the ß-tubulin gene has been attributed to the occurrence of resistance. In the present study, we investigated the presence of those SNPs in the ß-tubulin isotype-1 gene in different isolates of Parascaris in horse. Also, the mitochondrial (mt) and ribosomal genes were sequenced for species confirmation of the isolates. The analysis of sequences inferred from COII gene confirmed that those isolates were P. equorum. The distance between mt genes obtained here and several ascarid species in equids and other hosts suggests the need for the combination of more genetic data with morphologic and other diagnostic measures. The analysis on ß-tubulin isotype-1 gene revealed no resistance-related SNPs or substitutions at the expected codon positions and selection pressure with BZs has not occurred for Parascaris worms. Although the molecular data showed the susceptibility of Parascaris isolates against BZs, other mechanisms of resistance should be also investigated to confirm the validity of molecular results.

Therapeutic effect of simvastatin on DMBA-induced breast cancer in mice.

Preclinical studies have shown positive effects of statins against specific cancers. This study aimed to determine the therapeutic effect of simvastatin in 12-dimethylbenz(a)anthracene (DMBA)-induced breast cancer. Female albino mice were divided into two groups, with or without DMBA administration. After tumor appearance, DMBA-treated group was further divided into four groups (D1-D4) as control (D1), treated with simvastatin at 80 and 40 mg/kg/day, orally (D2 and D3) and tamoxifen (50 mg/kg/day, orally) treated group (D4). After 4 weeks, animals were sacrificed, serum samples were collected and tumors were dissected for histopathological study and determination of selected parameters. The tumor marker carcinoma antigen 15-3 (CA15-3), oxidative stress parameters and prostaglandin E2 (PGE2) levels were analyzed in serum and tumors in experimental groups. Tamoxifen and high dose of simvastatin improved parameters of mammary carcinogenesis including mean tumor volume, body weight and percent of mortality as compared to mice with breast tumors without treatment (D1). Additionally, simvastatin usage increased total antioxidant capacity (TAC) level, paraoxonase 1 (PON1) activity in serum and decreased total oxidant status (TOS) and malondialdehyde (MDA) levels in tumors similar to tamoxifen. No significant decrease was found in serum CA 15-3 and tumor PGE2 levels in simvastatin and tamoxifen treated groups as compared to D1 group. These data suggest that simvastatin has anticancer effects which are relatively similar to that of tamoxifen in an animal model of breast cancer.