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Lotmaria passim (Kinetoplastea) is considered the most prevalent as well as the most virulent trypanosomatid associated to the European honey bee Apis mellifera. We used qPCR to screen for the presence of this parasite in 57 samples from ten Argentinian provinces, and were able to detect its presence throughout most of the country with 41% of the samples testing positive. In a retrospective analysis, we detected L. passim in 73% of honey bee samples from 2006 showing that this flagellate has been widely present in Argentina for at least ~15 years. Additionally, three primer sets for L. passim detection were compared, with the pair that produced smallest PCR product having the best detection capability. Finally, we also found L. passim DNA in 100% (n = 6) of samples of the mite Varroa destructor. The role of this ectoparasite in the lifecycle of Lotmaria , if any, remains unrevealed. Unlabelled Image • The trypanosomatid Lotmaria passim is widely present in Argentina since 2006. • Forty-one percent of the analyzed honey bees were Lotmaria passim -positive. • All analyzed Varroa destructor individuals harbored DNA of Lotmaria passim. [ABSTRACT FROM AUTHOR]

• Leptospira spp. DNA by qPCR in aborted bovine fetuses was determined. • Concordance coefficient between qPCR and immuno-fluorescence antibody test was 0.44. • qPCR detection rate (32.4 %) in fetal tissues was higher compared with DFAT (11.8 %). • Ten out of eleven qPCR-positive fetuses had compatible histopathological findings. • qPCR was a rapid and sensitive method for detection of Leptospira in bovine fetuses. Leptospirosis is a disease with major economic impact on livestock industry. The objective of this work was to determine the presence of Leptospira spp. DNA by qPCR in bovine fetuses with presumptive diagnosis of leptospirosis as the cause of abortion. Leptospira spp. DNA was detected by qPCR in 11 out of 34 fetuses. These specimens (10/11) had histopathological findings in hepatic and/or renal tissues compatible with leptospirosis. qPCR detection rate (32.4 %) was higher compared with direct immuno-fluorescence antibody test (DFAT) (11.8 %). The concordance coefficient between both techniques was 0.44. qPCR is a rapid and sensitive technique for the diagnosis of leptospirosis and improved the detection rate in fetal tissues compared with DFAT. Implementation of molecular techniques may increase the accurate detection of leptospirosis as a cause of bovine abortion allowing the application of rapid therapeutic and prophylactics measures in order to reduce the impact of this zoonotic disease. [ABSTRACT FROM AUTHOR]

Zinc (Zn) is a vital trace element for the body and its bioavailability influences numerous reproductive events. However, the mechanisms that regulate Zn homeostasis in the cumulus-oocyte complex (COC) are yet to be elucidated. The aim of this study was to investigate the role of estradiol 17-beta (E 2), FSH and LH in Zn homeostasis regulation in bovine COC matured in vitro and Zn transporters gene expression. For this purpose, intracellular Zn levels in oocytes and cumulus cells (CC) were assessed using a Zn-specific fluorescent indicator. In addition, gene expression and sequencing of six Zn transporters (Slc39a6, Slc39a8, Slc39a14, Slc30a3, Slc30a7 and Slc30a9) were assessed. Our results demonstrated that the simultaneous presence of E 2 , FSH, and LH during oocyte maturation altered intracellular zinc levels and transporters expression in both oocytes and CC. Transporter's gene expression was different in oocytes and CC, possibly due to cell-specific changes in Zn levels during maturation. The interaction effects of Zn with hormonal treatments influenced the results. This study emphasizes that Slc39a6 is highly sensitive to hormone induction. Overall, the hormonal modulation of Zn homeostasis in the COC was evidenced. Also, a preponderant role of FSH as a modulator of Zn intracellular levels and transporter gene expression is suggested. • Zinc availability is beneficial for follicular growth, oocyte maturation, fertilization, and embryo development. • Reproductive hormones affected Zn homeostasis and gene expression of Zn transporters during IVM of bovine COC. • The gene expression of Zn transporters in oocytes and cumulus cells were different. • The role of FSH in the modulation of intracellular Zn levels and transporter gene expression is evident. • The Zn transporter Slc39a6 is highly sensitive to hormonal regulation. [ABSTRACT FROM AUTHOR]

Bovine alphaherpesvirus type 1 (BoAHV-1) is associated with respiratory and reproductive syndromes. Until present the immunologic mechanisms involved in BoAHV-1 abortion are partially known. We studied key elements of the innate immune response in the placentas and fetal lungs from cattle experimentally-inoculated with BoAHV-1. These tissues were analyzed by histopathology. Furthermore, virus identification was performed by qPCR and the expression of the inflammatory cytokines such as tumor necrosis factor-alpha, interleukin 1-alpha and inflammatory mediators like inducible nitric oxide synthase and cyclooxeganse-2 was evaluated by immunohistochemistry. The viral transplacental infection was confirmed by the detection of BoAHV-1 by qPCR in the placenta and fetal organs, which revealed mild inflammatory lesions. Inducible nitric oxide synthase immunolabelling was high in the lungs of infected fetuses and placentas, as well as for tumor necrosis factor-alpha in the pulmonary parenchyma and cyclooxeganse-2 in fetal annexes. However, the expression of interleukin 1-alpha was weak in these organs. To our knowledge, this is the first study that provides strong evidence of an early immune response to BoAHV-1 infection in the conceptus. Advances in the knowledge of the complex immunological interactions at the feto-maternal unit during BoAHV-1 infection are needed to clarify the pathogenesis of abortion. • BoAHV-1 transplacental infection causes an early immune response at the feto-maternal unit. • BoAHV-1 induces protein expression of iNOS in fetal lung and placenta. • BoAHV-1 induces protein expression of TNFα in fetal lungs and COX-2 in the placenta. • The expression of pro-inflammatory mediators in the placenta by BoAHV-1 might trigger the abortion or control the infection. [ABSTRACT FROM AUTHOR]

Objective: To study expression of dysadherin in human testis, epididymis, and spermatozoa. Design: Prospective study. Setting: Basic research laboratory. Patient(s): Testis, epididymis, and testicular spermatozoa from patients under treatment and semen from volunteer donors. Intervention(s): Reverse transcription–polymerase chain reaction, immunohistochemistry, immunocytochemistry, and Western immunoblotting. Main Outcome Measure(s): Dysadherin messenger RNA (mRNA) analysis in testis, epididymis, and ejaculated spermatozoa, immunohistochemistry of both tissues, Western immunoblotting of tissue/cell extracts, and immunocytochemistry of spermatozoa. Result(s): Dysadherin mRNA was found in testis, epididymis, and ejaculated spermatozoa. Whereas testis and spermatozoa exhibited a distinctive 91-kDa protein form, the epididymis showed a 50-kDa moiety, also found in MDA-MB-231 breast cancer cells. Nucleotide sequence analysis revealed >99% homology between testicular and somatic cell mRNA, suggesting differential protein glycosylation. Dysadherin was immunodetected in round spermatids and testicular/ejaculated spermatozoa. It localizes to the acrosomal region and flagellum and colocalized with E-cadherin in the head and with the Na+,K+-ATPase α4 subunit in the flagellum. Conclusion(s): This is the first report on expression of dysadherin in the male gonad and in spermatozoa. Its colocalization with E-cadherin and Na+,K+-ATPase leads us to postulate a role for dysadherin as a modulator of sperm function. [Copyright &y& Elsevier]