There is little information on human Q fever in Iran and other Middle East countries. The aim of this study was to determine apparent Q fever seropositivity among febrile patients with suspected brucellosis in southeast Iran. Coxiella burnetii phases I and II specific IgG antibodies were measured by a commercial enzyme-linked immunosorbent assay (ELISA) in sera from 75 febrile patients. Phase I antibodies were detected in 18 subjects (24%) and phase II antibodies in 27 subjects (36%). This is the first report of human Q fever seropositivity in Iran after 3 decades and demonstrated a high prevalence of C. burnetii exposure in the sampled febrile patients.

Clostridium perfringens type D infects ruminants and causes the enterotoxemia disease by ε-toxin. A mutated ε-toxin gene lacking toxicity was designed, synthesized, and cloned into the pT1NX vector and electroporated into Lactobacillus casei competent cells to yield LC-pT1NX-ε recombinant strain. BALB/c mice, immunized orally with this strain, highly induced mucosal, humoral, and cell-mediated immune responses and developed a protection against 200 MLD/ml of the activated ε-toxin. This study showed that the LC-pT1NX-ε could be a promising vaccine candidate against the enterotoxemia disease.

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Background: There are several protocols to extract DNA from Lactobacillus spp. In the case of L. casei it is harder because of its especial and thick cell wall. Objectives: In this study, nine DNA extraction protocols (by lysozyme treatment) were evaluated and compared in two categories (traditional and kit-based protocols) and an improved method was presented. Materials and Methods: DNA quantity and quality was determined by spectrophotometry, agarose gel electrophoresis and polymerase chain reaction (PCR). Results:The results revealed that the yield of extracted DNA differed by each protocol (5.8 -17.1 μg/100 μL), but provided appropriate DNA for PCR amplification. The modified protocol offered the best total DNA extraction method when both quality (DNA purity; 1.54 μg) and quantity (DNA yield; 17.1 μg) were considered. Conclusions: We suggest this protocol for effective and inexpensive DNA isolation from L. casei for downstream biological processes such as PCR.

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Background Epsilon toxin (ETX) causes a commonly fatal enterotoxemia in domestic animals. Also, ETX causes serious economic losses to animal husbandry. In this study, we selected several clones against ETX using repertoires displayed on filamentous phage. Anti-ETX specific clones were enriched by binding to immobilized antigen, followed by elution and re-propagation of phage. After multiple rounds of binding selection, ELISA analysis showed that most isolated clones had high affinity and specificity for ETX. Results Two recombinant monoclonal antibodies against ETX were isolated by phage display technology. B1 phage VH antibody isolated from DAb library and G2 soluble scFv antibody isolated from Tomlinson I + J libraries have been applied as the capture and detection antibodies for developing an ETX sandwich ELISA test, respectively. Conclusions Designed ETX sandwich ELISA could be a valuable tool for quantitative detection of ETX in inactivated commercial vaccines against enterotoxemia. Graphical abstract

Background: Escherichia coli can be categorized into four major phylogenetic groups (A, B1, B2, and D) based on presence or absence of three markers including two genes (chuA and yjaA) and an anonymous DNA fragment designated TSPE4.C2. Also, these groups are divided into seven phylogenetic subgroups A 0 , A 1 , B1, B2 2 , B2 3 , D 1 , and D 2 . Objectives: This study aimed to determine the distribution pattern of phylogenetic groups in Shiga Toxin-Producing Escherichia coli (STEC) and Enteropathogenic Escherichia coli (EPEC) isolated from asymptomatic goats in Kerman city, Iran. Materials and Methods: Two hundred and fifty fecal samples were obtained from healthy goats. All isolates were subjected to detection of phylogenetic markers chuA, yjaA and DNA fragment TspE4.C2 and virulence genes stx1, stx2 and eae. Results: In summary, among all isolates phylo-group B1 was the most prevalent (57.6%) and other phylo-groups were A 1 (20.4%), A 0 (18.4%), D 1 (2.8%), and B2 2 (0.8%). There was no isolate in B2 3 and D 2 subgroups. Fifty samples (20%) possessed at least one of the tested virulence genes: stx1 (12%), stx1/stx2 (4%), eae (2.8%), stx1/eae (0.8%), and stx2 (0.4%). Thus, 41 (16.4%) STEC, 7 (2.8%) EPEC, and 2 (0.8%) Enterohemorrhagic Escherichia coli (EHEC) strains were isolated and allocated into four phylogenetic subgroups A 0 (16%), A 1 (12%), B1 (68%), and D 1 (4%). Conclusions: Based on 250 fecal samples obtained from goats in industrial slaughterhouse of Kerman City, goats may be a potential reservoir of STEC in Kerman and B1 followed by A are the most prevalent phylogenetic groups among STEC and non-STEC isolates in this study.