BACKGROUND: Various flea species have already been reported from dogs, among which the most important ones include Ct. felis, Ct. canis, and P. irritans. Fleas can cause annoyance in dogs and human and transmit a variety of bacterial, fungal, and viral agents to the host. In addition, they could function as an intermediate host of Dipylidium caninum and Hymenolepis diminuta.  OBJECTIVES: Due to the lack of molecular species-associated identification data, we conducted the current study to differentiate Ct. felis and Ct. canis with molecular assay. METHODS: In the present study, 605 fleas were primarily collected from the dogs referred to Tehran Veterinary Faculty hospital. Subsequently, the flea species were identified under a microscope with morphological keys. Afterwards, COX1 genes of Ct. felis and Ct. canis were amplified via PCR and the locus was finally compared utilizing RFLP and sequencing. RESULTS: Totally, 605 fleas were isolated from 20 dogs. In morphological studies, three species were identified: Ctenocephalides felis, Ctenocephalides canis, Pulex irritans. Pulex irritans had the highest frequency (61.8 %). In molecular study, 552 bp fragment of COX1 gene in two species was amplified and seen on agarose gel. After sequencing, it was seen that two species sequences in COX1 locus had a similarity of 99 % and all of them depended on Ct. canis. In PCR-RFLP, in which Taq1 enzyme was used for differentiation of two species, the same result was obtained. CONCLUSIONS: Even though these two species of dog flea are distinct morphologically, their molecular differentiation using COX1 genes was not successful.

BACKGROUND: Dactylogyrus is one of the most common external parasites on                the gills of Cyprind fish. These parasites are highly host specific and many species only have a specific host. OBJECTIVES: Since there are reports of silver carp specific Dactylogyrus species isolated from big head carp and vice versa, the investigation of Dactylogyrids have been done in these two fish species. METHODS: 81 silver carp and 82 big head carp were caught from 10 fish farms in Guilan province and after preparing wet mounts of body surface Dactylogyrus parasites divided and fixed by glycerin jelly. In order to perform morphometric assessments on captured images, Image J software was used for 7 point to point distances. Drawing of parasites was done by drawing tube and then compared by identification keys and parasites identified. For molecular investigation the genomic DNA was extracted from one parasite specimen and 28S rDNA region of Dactylogyrus specimens were amplified by related primers in PCR. RESULTS: Sequences were deposited in GenBank with accession numbers MG825611 and MG825765 respectively for D. hypophthalmichthys and D. suchengtaii isolated from Hypophthalmichthys molitrix, and also MH023397 and MH023399 respectively for D. aristichthys and D. nobilis isolated from Hypophthalmichthys nobilis. The phylogenetic tree shows the genetic affinity of isolated parasites from these two fish. CONCLUSIONS: It seems hybrid fish are sometimes produced accidentally in fish reproduction centers of Iran. Racial impurity of silver carp and big head carp is not only the reason of poorer breeding efficiency in fish farms but also these hybrid fish are hosts of more parasitic species.

Two hundred ninety-six Eschericbia coli isolates from feces or intestines of calves with diarrhea were hybridized with 7 gene probes. One probe (the eae probe) was derived from the eae gene coding for a protein involved in the effacement of the enterocyte microvilli by the group of bacteria called attaching and effacing E coli (AEEC), and 2 probes were derived from genes coding for the Shiga-like toxins (SLT) 1 and 2 produced by the verocytotoxic E coli (VTEC). The other 4 probes were derived from DNA sequences associated with the adhesive properties of enteroadherent E coli (EAEC) to cultured cells (the EAF probe for the localized adherence pattern, probes F1845 and AIDA-1 for the diffuse adherence pattern, and the Agg probe for the aggregative adherence pattern). Hybridization results for the eae probe were in agreement, for all but 1 of the 8 isolates, with previously published phenotypic results of microvilli effacement. The latter was previously reported as effacing the microvilli of calf enterocytes, but was eae probe-negative. Two classes of isolates hybridized with the eae probe. Members of a first class (60 isolates) additionally produced a positive signal with 1 or both of the SLT probes (VTEC-AEEC isolates). Isolates hy- bridizing with the eae and the SLT1 probes were the most frequent: 56 isolates (ie, 93% of all VTEC-AEEC). Members of the second class (10 isolates) failed to hybridize with either SLT probe (non-VTEC-AEEC isolates). Most isolates of these 2 classes belong to only 4 serogroups: O5, O26, O111, and O118. In addition to these 2 AEEC classes, a VTEC class (20 isolates) was observed. Such isolates were positive with 1 or both SLT probes, but were negative with the eae probe. All but 1 isolate belonged to serogroups not found among the AEEC isolates. Only 7 of all AEEC and VTEC isolates were positive with the EAF, the F1845, or the AIDA-1 probe, and none were positive with the Agg probe. On the other hand, 32 non-VTEC, non-AEEC isolates were po.

Colony hybridizations with DNA probes for 3 heat-stable (STaP, STaH, and STb) enterotoxins and 1 heat-labile (LT) enterotoxin and for 4 adhesins (K99, F41, K88, 987P) were performed on 870 Escherichia coli isolates to determine pathotypes prevalent among enterotoxigenic E coli (ETEC) isolated from cattle in Belgium. One hundred thirty-two E coli isolates (15.2%) hybridized with probes STaP, K99, and/or F41. The 5 other probes were not hybridized by E coli isolates. Therefore, only STaP enterotoxin and K99 and F41 adhesins were virulence factors of ETEC isolated from cattle. Two major pathotypes accounted for 95% of the ETEC: STaP+K99+F41+ (67.4%) and STaP+K99+ (27.3%). The last 5% of probe-positive isolates had STaP+, STaP+F41+, or K99+F41+ minor pathotypes. Of 12 American ETEC isolates also assayed, 7 were positive with STb and/or 987P probes (pathotypes STaP+STb+,STaP+ 987P+, or STaP+STb+987P+) and may be porcine- rather than bovine-specific enteropathogens. The remaining 5 American ETEC isolates belonged to 3 minor pathotypes (STaP+,STaP+F41+, and K99+F41+) also found among Belgian E coli isolates. Such isolates may be derivatives of STaP+K99+F41+ or STaP+K99+ ETEC after in vivo or in vitro loss of virulence genes and/or non-ETEC isolates, which have acquired virulence genes by in vivo transfer.

The aim of this work was the validation of a rapid real-time PCR assay based on TaqMan technology for the unequivocal identification of infectious canine hepatitis (ICH) virus, to be used directly on DNA purified from blood specimens. A real-time PCR system targeting at the E3 ORFA gene sequence of canine adenovirus type 1 was optimized and validated through comparative analysis of samples using conventional PCR system. The real-time PCR assay based on TaqMan technology could disclose 23 (37.7%) out of 61 samples as PCR positive. In contrast, 18 (29.5%) samples were found PCR positive when conventional PCR was applied on these samples.

Sal enteritidis thin fimbriae, SEF14, were found to be restricted to the predominantly poultry-associated members of the Salmonella serogroup D1 that are considered as the important pathogens in poultry industry. SefA together with sefB and sdfC encode the proteins involved in SEF14 biosynthesis. In order to develop the rapid and specific detection methods for Salmonella serogroup D1, a PCR technique for the am;lification of sefA gene was established, and its specificity and sensitivity were investigated with various microorganisms. The bacterial genomic DNA was extracted by colony-picking and rapid boiled-lysate technique. In comparison of SefI and SefII primers used in the PCR. SefI primer for sefA gene of 513bp showed higher specificity than that of SefII. The established PCR was s sensitive as to detect 1pg of Sal enteritidis DNA. When 73 strains in 28 genera including the reference strains and the field isolates of various Salmonella serotypes, Bacillus subtilis, Bordetella bronchiseptica, E coli, Listeria spp., Micrococcus luteus, Rhodococcus equi, Staphylococcus spp., Streptococcus spp., Vibrio parahemolyticus, Yersinia spp. were studied, the established PCR yielded specifically positive results with only Salmonella serogroup D1. The results suggested that the PCR for sefA gene could be a potential candidate among the specific detection methods for Salmonella serogroup D1.

The melanocortin 1 receptor (MC1R) plays an important role in regulation of melanin pigment synthesis within mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat color variations within several mammalian species including cattle. To develope a rapid and accurate method for the identification of Hanwoo meat, we performed a single nucleotide polymorphism (SNP) analysis in Melanocortin 1 receptor (MC1R) gene using TaqMan∨® MGB probe-based real-time PCR. Two specific probes (one for Hanwoo and the other for Holstein and Black angus) were designed.