Transferrins Reduce Replication of Chlamydia suis in McCoy Cells

Chlamydia suis (C. suis) resides in the intestines of pigs and tetracycline-resistant strains are emerging worldwide. Intestinal infections are often subclinical. However, the gut is regarded as a C. suis reservoir and clinical infections have been associated with enteritis, conjunctivitis, pneumonia and reproductive failure. C. suis was found in boar semen and venereal transmission occurred. We studied the anti-Chlamydia suis activity of ovotransferrin (ovoTF) and bovine lactoferrin (bLF). Pre-incubation of C. suis with bLF or ovoTF had no significant effect on overall chlamydia replication (mean fluorescence area) in McCoy cells. The addition of ovoTF to the culture medium had no effect on bacterial replication, but the addition of 0.5 or 5 mg/mL of bLF significantly reduced the inclusion size by 17% and 15% respectively. Egg components are used for cryopreservation of boar semen. When inoculating an ovoTF-containing and Chlamydia suis-spiked semen sample in McCoy cells, a significant reduction in inclusion number (by 7%) and overall replication (by 11%) was observed. Thus, we showed that transferrins possess anti-chlamydial activity. Moreover, ovoTF addition to semen extenders might reduce C. suis venereal transmission. Further research is needed to unravel the mechanisms behind the observations and to enhance the effect of transferrins on C. suis.

Genital Brucella suis Biovar 2 Infection of Wild Boar (Sus scrofa) Hunted in Tuscany (Italy)

Brucellosis is a zoonosis caused by different Brucella species. Wild boar (Sus scrofa) could be infected by some species and represents an important reservoir, especially for B. suis biovar 2. This study aimed to investigate the prevalence of Brucella spp. by serological and molecular assays in wild boar hunted in Tuscany (Italy) during two hunting seasons. From 287 animals, sera, lymph nodes, livers, spleens, and reproductive system organs were collected. Within sera, 16 (5.74%) were positive to both rose bengal test (RBT) and complement fixation test (CFT), with titres ranging from 1:4 to 1:16 (corresponding to 20 and 80 ICFTU/mL, respectively). Brucella spp. DNA was detected in four lymph nodes (1.40%), five epididymides (1.74%), and one fetus pool (2.22%). All positive PCR samples belonged to Brucella suis biovar 2. The results of this investigation confirmed that wild boar represents a host for B.suis biovar. 2 and plays an important role in the epidemiology of brucellosis in central Italy. Additionally, epididymis localization confirms the possible venereal transmission.

Leishmania infantum in the reproductive organs of dogs

ABSTRACT: Leishmania infantum causes canine leishmaniasis. Using parasitological and molecular analyses, we identified L. infantum in the reproductive organs of male and female dogs. Using histochemistry, immunohistochemistry, and PCR, we examined tissue samples from the reproductive organs of 8 male dogs and 16 female dogs diagnosed with leishmaniasis. Despite the absence of macroscopic or microscopic lesions in these organs, we observed L. infantum amastigotes in tissue samples from the testis and the uterus. PCR and sequencing of these tissues revealed sequences that matched 100% with L. infantum DNA available at GenBank. The presence of L. infantum amastigotes and DNA in testicular and uterine tissue samples suggested that these organs can harbor the parasite without associated macroscopic or microscopic lesions, and this can be especially important in the vertical and venereal transmission of leishmaniasis in dogs.

Presence of pathogenic Leptospira spp. in the reproductive system and fetuses of wild boars (Sus scrofa) in Italy

Leptospirosis is a re-emerging and globally spread zoonosis caused by pathogenic genomospecies of Leptospira. Wild boar (Sus scrofa) are an important Leptospira host and are increasing in population all over Europe. The aim of this investigation was to evaluate Leptospira spp. infection in the reproductive systems of wild boar hunted in two Italian regions: Tuscany and Sardinia. From 231 animals, reproductive system tissue samples (testicles, epididymides, uteri) as well as placentas and fetuses were collected. Bacteriological examination and Real-Time PCR were performed to detect pathogenic Leptospira (lipL32 gene). Leptospires were isolated from the testicles and epididymides of one adult and two subadult wild boar. Four isolates from the two subadult males were identified as Leptospira interrogans serogroup Australis by MLST, whereas Leptospira kirschneri serogroup Grippotyphosa was identified from the adult testicles and epididymis. Using Real-Time PCR, 70 samples were positive: 22 testicles (23.16%) and 22 epididymides (23.16%), 10 uteri (7.35%), 3 placentas (6.66%), and 13 fetuses (28.88%). Amplification of the rrs2 gene identified L. interrogans and L. kirschneri species. The results from this investigation confirmed that wild boar represent a potential source of pathogenic Leptospira spp. Isolation of Leptospira serogroups Australis and Grippotyphosa from the male reproductive system and the positive Real-Time PCR results from both male and female samples could suggest venereal transmission, as already demonstrated in pigs. Furthermore, placentas and fetuses were positive for the lipL32 target, and this finding may be related to a possible vertical transmission of pathogenic Leptospira.

Prevalence of Coxiella burnetii in German sheep flocks and evaluation of a novel approach to detect an infection via preputial swabs at herd-level

A prevalence study was conducted on German sheep flocks including goats if they cohabitated with sheep. In addition, a novel approach was applied to identify an infection at the herd-level before lambing season with preputial swabs, suspecting venereal transmission and ensuing colonisation of preputial mucosa with Coxiella (C.) burnetii. Blood samples and genital swabs were collected from breeding males and females after the mating season and were analysed by enzyme-linked immunosorbent assay (ELISA) and quantitative polymerase chain reaction (qPCR) respectively. In total, 3367 animals were sampled across 71 flocks. The true herd-level prevalence adjusted for misclassification probabilities of the applied diagnostic tests using the Rogan-Gladen estimator for the prevalence estimate and a formula by Lang and Reiczigel (2014) for the confidence limits, ranged between 31.3% and 33% (95% confidence interval [95% CI] 17.3–45.5) detected by the ELISA and/or qPCR. Overall 26–36.6% (95% CI 13–56.8) were detected by ELISA, 13.9% (95% CI 4.5–23.2) by the qPCR and 7.9–11.2% (95% CI 0.08–22.3) by both tests simultaneously. The range of results is due to data obtained from literature with different specifications for test quality for ELISA. Among eight farms with females shedding C. burnetii, three farms (37.5%) could also be identified by preputial swabs from breeding sires. This indicates less reliability of preputial swabs if used as a single diagnostic tool to detect C. burnetii infection at the herd-level.

Epidemiology and Control of Congo Fever in Sacrificial Animals of Pakistan

The cases and deaths due to Crimean-Congo haemorrhagic fever (CCHF) [49] virus commonly known as Congo virus (fatality rate 15%) have been reported throughout Pakistan from the last five years especially during religious occasion, Eid-ul-Azha. The annual increase in death rates due to CCHF demonstrate the importance of awareness of Congo fever at academia as well as public level. The symptoms of Congo fever which appear one to nine days after tick bite, include sudden high fever, muscle aches, abdominal pain, headache, dizziness, sore eyes, jaundice, mood swings, confusion, aggression, and sensitivity to light. The other signs include sore throat, joint pain, vomiting, diarrhea, hemorrhages, and bleeding from skin and large intestine. The Infection has been reported in many species of wild as well as domestic animals including hares, cattle, sheep, goats, dogs, mice and hedgehogs. At least 31 species of Hyalomma, Boophilus, Rhipicephalus, Dermacentor (Ixodidae: hard ticks) act as vector of CCHF in which transovarial, transstadial and venereal transmission occurs. The virus attacks the immune system of the host and influences the immune cells. The Congo fever virus can be isolated from blood, plasma and many body tissues (kidneys, liver, spleen, lungs, brain and bone marrow). Mice inoculation, enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR) can be used for detection of the infection. Furthermore, IgM and IgG antibodies against CCHFV can also be detected and quantified. Education of general public, tick control with acaricides, use of anti-CCHFV immunoglobulin, usage of approved repellents to prevent tick bites, wearing neutral-coloured garments, application of a permethrin spray to the clothing, avoiding tall grasses and shrubs, applying sunscreen, avoiding direct contact with the blood or tissues of animals are the factors for successful prevention of the infection.

The Potency of Polygamy Behavior in Aedes aegypti Mosquitoes by Venereal Transmission Dengue Virus

Male Aedes aegypti mosquito has been considered to not have any important role in transmitting dengue virus (DENV). The purpose of this study is to prove that male Ae. aegypti mosquito does have an important role in transmitting DENV 3 through venereal transmission with their potency of its polygamy behavior. The data collection was done using colonization method and intrathoracal injection. The presence of DENV 3 on male and female mosquitoes was proven by RT-PCR method (profile of DNA band specifically on 511 bp) and serotyping PCR (290 bp). The data were analyzed using univariate analysis followed by bivariate analysis with parametric test ANOVA. The result of the study demonstrated that male Ae. aegypti mosquitoes do have an important role in transmitting DENV 3 through venereal transmission with the potency of their polygamy behavior. There was no significant difference between the polygamy behaviors of Ae. aegypti male mosquito infected by DENV and the non-infectious Ae. aegypti male mosquito.