Tularemia and Plague Survey in Rodents in Earthquake Zones in Southeastern Iran

OBJECTIVES: Earthquakes are one the most common natural disasters that lead to increased mortality and morbidity from transmissible diseases, partially because the rodents displaced by an earthquake can lead to an increased rate of disease transmission. The aim of this study was to evaluate the prevalence of plague and tularemia in rodents in the earthquake zones in southeastern Iran. METHODS: In April 2013, a research team was dispatched to explore the possible presence of diseases in rodents displaced by a recent earthquake magnitude 7.7 around the cities of Khash and Saravan in Sistan and Baluchestan Province. Rodents were trapped near and in the earthquake zone, in a location where an outbreak of tularemia was reported in 2007. Rodent serums were tested for a serological survey using an enzyme-linked immunosorbent assay. RESULTS: In the 13 areas that were studied, nine rodents were caught over a total of 200 trap-days. Forty-eight fleas and 10 ticks were obtained from the rodents. The ticks were from the Hyalomma genus and the fleas were from the Xenopsylla genus. All the trapped rodents were Tatera indica. Serological results were negative for plague, but the serum agglutination test was positive for tularemia in one of the rodents. Tatera indica has never been previously documented to be involved in the transmission of tularemia. CONCLUSIONS: No evidence of the plague cycle was found in the rodents of the area, but evidence was found of tularemia infection in rodents, as demonstrated by a positive serological test for tularemia in one rodent

Establishment of a Mycoplasma hyorhinis challenge model in 5-week-old piglets

IntroductionMycoplasma hyorhinis is an emerging swine pathogen with high prevalence worldwide. The main lesions caused are arthritis and polyserositis, and the clinical manifestation of the disease may result in significant economic losses due to decreased weight gain and enhanced medical costs. We aimed to compare two challenge routes to induce M. hyorhinis infection using the same clinical isolate.MethodsFive-week-old, Choice hybrid pigs were inoculated on 2 consecutive days by intravenous route (Group IV-IV) or by intravenous and intraperitoneal routes (Group IV-IP). Mock-infected animals were used as control (control group). After the challenge, the clinical signs were recorded for 28 days, after which the animals were euthanized. Gross pathological and histopathological examinations, PCR detection, isolation, and genotyping of the re-isolated Mycoplasma sp. and culture of bacteria other than Mycoplasma sp. were carried out. The ELISA test was used to detect anti-M. hyorhinis immunoglobulins in the sera of all animals.ResultsPericarditis and polyarthritis were observed in both challenge groups; however, the serositis was more severe in Group IV-IV. Statistically significant differences were detected between the challenged groups and the control group regarding the average daily weight gain, pathological scores, and ELISA titers. Additionally, histopathological scores in Group IV-IV differed significantly from the scores in the control group. All re-isolated strains were the same or a close genetic variant of the original challenge strain.DiscussionOur results indicate that both challenge routes are suitable for modeling the disease. However, due to the evoked more severe pathological lesions and the application being similar to the hypothesized natural route of infection in Group IV-IV, the two-dose intravenous challenge is recommended by the authors to induce serositis and arthritis associated with M. hyorhinis infection

Development of molecular biological tools for the rapid determination of antibiotic susceptibility of Mycoplasma hyopneumoniae isolates

Mycoplasma hyopneumoniae is the etiologic agent of porcine enzootic pneumonia, a contagious respiratory disease, causing significant economic losses worldwide. Antibiotic treatment is commonly utilised in the pig industry to control M. hyopneumoniae infection. Since the conventional antibiotic susceptibility test is time-consuming, taking up to weeks’ period, antibiotics are usually empirically chosen. Certain single nucleotide polymorphisms in the parC (C239A/T, G250A) and gyrA (G242C, C247 T, A260 G) genes show correlation with decreased fluoroquinolone susceptibility by the change of the target site. Furthermore, the nucleotide alteration A2059 G in the 23S rRNA sequence correlates with significantly decreased macrolide and lincosamide susceptibility of M. hyopneumoniae. Mismatch amplification mutation assays (MAMA) and high resolution melt (HRM) analysis, capable to detect the mentioned resistance markers, were developed in the present study, in order to provide susceptibility data in a considerably shorter time than the conventional methods. The results of the MAMA and HRM assays were congruent with the results of the conventional antibiotic susceptibility method of the tested M. hyopneumoniae field isolates. The sensitivity of the MAMAs was 103-104 copy numbers, while that of the HRM assay was 105-106 copy numbers. To the best of our knowledge this was the first time that MAMA and HRM assays were developed for the rapid detection of decreased fluoroquinolone, macrolide or lincosamide susceptibility in M. hyopneumoniae strains

Screening of bat faeces for arthropod-borne apicomplexan protozoa: Babesia canis and Besnoitia besnoiti-like sequences from Chiroptera

Background : 45 Microbats (Chiroptera: Microchiropte ra) are among the most eco - epidemiologically important 46 mammals, owing to their presence in human settlements and ani mal keeping facilities . 47 Roosting of bats in buildings may bring pathogens of veterinary - medical importance into the 48 environment of domestic animals and humans. In this context bats have long been studied as 49 carriers of various pathogen groups. However, despite their close association with arthropods 50 (both in their f oo d and as their ectoparasites), only a few molecular surveys have been 51 publish ed on their role as carriers of vector - borne protozoa. The aim of the present study was 52 to compensate for this scarcity of information. 53 Findings : 54 Altogether 221 (mostly individual) bat faecal samples were collected in Hungary and the 55 Netherlands. The DNA w as extracted , and analysed with PCR and sequencing for the 56 presence of arthropod - borne apicomplexan protozoa. Babesia canis canis (with 99 - 100% 57 homology) was identified in five samples, all from Hungary. Because it was excluded with an 58 Ixodidae - specific PC R that the relevant bats consumed ticks, these sequences derive either 59 from insect carriers of Ba. canis , or from the infection of bats. In one bat faecal sample from 60 the Netherlands a sequence having the highest (99%) homology to Besnoitia besnoiti was 61 am plified. 62 Conclusions : 63 The se findings suggest that some aspects of the epidemiology of canine babesiosis are 64 underestimated or unknown, i.e. the potential role of insect - borne mechanical transmission 65 and/or the susceptibility of bats to Ba. canis . In addit ion, b ats need to be added to future 66 studies in the quest for the final host of Be. besnoiti

Screening of Hungarian cattle herds for seropositivity to Mycoplasma bovis

A total of 860 serum samples collected at 86 cattle farms in different parts of Hungary were screened for the presence of antibodies to Mycoplasma bovis using an ELISA test with a recombinant M. bovis membrane protein as antigen. Antibodies to M. bovis were detected in sera collected on all farms, and no farms negative for M. bovis were found. In 88.38% of the herds more than 50% of the sampled animals were infected by M. bovis. A total of 82.91% of the animals had antibodies to M. bovis. The proportion of seropositive animals was higher in the older age groups, and a significant difference was seen in the level of seropositivity between young and older age groups. The results show that M. bovis infection is widespread on Hungarian dairy farms, and its prevalence has increased in the recent decade. The high infection rate of Hungarian cattle herds with M. bovis shows that special attention should be paid to evaluating the aetiological role of M. bovis in bovine respiratory disease complex (BRDC) cases because M. bovis has an immunosuppressive effect and can predispose cattle to other respiratory infections, too

Rapid, simple and cost-effective molecular method to differentiate the temperature sensitive (ts+) MS-H vaccine strain and wild-type Mycoplasma synoviae isolates

Mycoplasma synoviae infection in chickens and turkeys can cause respiratory disease, infectious synovitis and eggshell apex abnormality; thus it is an economically important pathogen. Control of M . synoviae infection comprises eradication, medication or vaccina- tion. The differentiation of the temperature sensitive (ts + ) MS-H vaccine strain from field iso- lates is crucial during vaccination programs. Melt-curve and agarose gel based mismatch amplification mutation assays (MAMA) are provided in the present study to distinguish between the ts + MS-H vaccine strain, its non-temperature sensitive re-isolates and wild- type M . synoviae isolates based on the single nucleotide polymorphisms at nt367 and nt629 of the obg gene. The two melt-MAMAs and the two agarose-MAMAs clearly distinguish the ts + MS-H vaccine strain genotype from its non-temperature sensitive re-isolate genotype and wild-type M . synoviae isolate genotype, and no cross-reactions with other Mycoplasma species infecting birds occur. The sensitivity of the melt-MAMAs and agarose-MAMAs was 10 3 and 10 4 copy numbers, respectively. The assays can be performed directly on clinical samples and they can be run simultaneously at the same annealing temperature. The assays can be performed in laboratories with limited facilities, using basic real-time PCR machine or conventional thermocycler coupled with agarose gel electrophoresis. The advantages of the described assays compared with previously used methods are simplicity, sufficient sensitivity, time and cost effectiveness and specificity

Antimicrobial susceptibility of Bacillus anthracis strains from Hungary

The susceptibility of 29 Bacillus anthracis strains, collected in Hungary between 1933 and 2014, was tested to 10 antibiotics with commercially available minimum inhibitory concentration (MIC) test strips. All strains were susceptible to amoxicillin, ciprofloxacin, clindamycin, doxycycline, gentamicin, penicillin, rifampicin, and vancomycin. Intermediate susceptibility to erythromycin and cefotaxime was detected in 17.2% (5/29) and 58.6% (17/29) of the strains, respectively. Correlations were not observed between the isolation date, location, host species, genotype, and antibiotic susceptibility profile of strains