Molekularbiologische Untersuchungen über Mycobacterium avium-intracellulare Komplex (MAIC) isoliert aus Schlachtschweinen und Wildtieren in Deutschland

Title and contents 1\. Introduction 2\. Review of literature 3\. Materials and methods 4\. Results 5\. Discussion 6\. Zusammenfassung 7\. Summary 8\. References 9\. Appendix I Abbreviations II List of solutions and buffers III List of equipments IV List of tables and figures 10\. AcknowledgementsIn a cross-sectional study multiple lymph nodes from 217 pigs, 174 from conventionally managed farms and 43 from an "eco" farm in different geographic areas of Germany slaughtered in four slaughterhouses were investigated for their mycobacterial burden. By carcass inspection 96 pigs were found to exhibit pathological lesions, 121 did not show macroscopic signs of mycobacterial infection. Lymph node specimens from different organs of all 217 pigs were investigated by Ziehl-Neelsen staining and cultivation. The species of isolates were identified by PCR. Staining showed sensitivity of 81,16% compared to culture of tissue on Löwenstein-Jensen or Stonebrink medium. From 32,05% of the conventionally managed pigs without pathological lesions mycobacteria could be isolated, compared to 60,46% of "eco" pigs. In 29.66% of the animals mycobacteria were found in multiple lymph nodes, in some cases also in muscle lymph nodes. In 83 out of 217 pigs M. avium subspecies hominissuis was isolated. High resolution genotyping using RFLP and PFGE showed a high degree of intrasubspecies differences, but no relationship between RFLP and PFGE clusters. PFGE of M. avium ssp. hominissuis isolates from pigs originating from different regions slaughtered in one of the slaughterhouses showed close clonal relationship to each other and to cattle isolates from the geographical area of the respective slaughterhouse found in the year 2000. This clonal group comprised about 45% of all M. avium ssp. hominissuis isolates, while RFLP yielded a very diffiuse dendrogramme. Therefore PFGE is considered to be the more suitable method for large scale epidemiological investigations for M. avium infections.Gegenstand der vorliegenden Arbeit ist die labordiagnostische Untersuchung des Vorkommens von Mycobacterium avium-intracellulare Komplex (MAIC) bei Schlachtschweinen. Die Probennahme wurde in Form eine Querschnittsstudie durchgeführt. Die Ergebnisse durch Ziehl-Neelsen Färbung zeigen relativ niedrige Sensitivität im Vergleich zu kultureller Technik. In dieser Studie ist Ogawa der beste Nährboden für die erste Isolierung. Die Versuche mit Löwenstein-Jensen (LJ) und Stonebrink (S) sollten parallel durchgeführt werden, weil einige Stämme nur auf LJ und/oder S wachsen. Die Schweine aus einem �Öko� Bestand zeigen etwa doppelt so hohe Prävalenzen (60.46%) wie die Schweine aus konventionellen Beständen (32.05%). Bei 29.66% waren mehr als ein Lymphknoten infiziert. Nach den Ergebnissen dieser Arbeit stellen mit Mykobakterien belastete Schweine, zumal da ein Prozentsatz der Tiere keine pathol.-anatomischen Veränderungen aufweist, eine mögliche Infektionsquelle für den Verbraucher. Anhand der Untersuchung mit PCR wurde Mycobacterium (M.) avium ssp. hominissuis am häufigsten identifiziert gefolgt von M.avium ssp.avium und selten M. intracellulare. Nach dem Ergebnis mit PFGE zeigen die M. avium ssp. hominissuis, die von Schlachtschweinen aus Essen (NI), Meiningen (TH) und Wildtiere aus Thüringen isoliert wurden, die ähnliche molekulare Charakteristika wie die Rinderstämme aus Cloppenburg in Jahr 2000. Dieses Ergebnis zeigt, dass PFGE für molekulare epidemiologische Studien über große geographische und zeitliche Räume besser geeignet ist

Assessment of knowledge, attitude and practices of people toward Japanese Encephalitis in endemic areas of Nepal

Japanese encephalitis is a serious vector borne viral zoonotic disease, particularly in Asia and Northern Australia. In Nepal, JE cases were reported in 63 out of 75 districts up until the year 2005. The aim of this study was to assess the level of knowledge, attitude and practices (KAP) toward JE and associated factors. A structured questionnaire was used to gather information on the KAP of people in endemic areas. Accordingly, 397 respondents demonstrated a desirable attitude, while half of the population exhibited desirable knowledge and practices. Age was found to be a significant factor for knowledge, wherein the senior population had better awareness of the vectors of JE. Moreover, education and occupation were found to be significant factors for KAP scores with an increase in level of education leading to an increase in desirable characteristics. This study determined that the characteristics of the surrounding environment were prominent factors. A significant number of people were not aware of the JE vaccine even though the government has made it available to most of the population of Nepal. Thus, more effective awareness programs should be implemented to increase the KAP of the population with regard to JE

Prevalence and demographic risk factors of Mycobacterium tuberculosis infections in captive Asian elephants (Elephas maximus) based on serological assays

To address putative TB statuses of elephants and to identify and quantify potential demographic risk factors for TB, three ELISAs specific for different mycobacterial antigens (ESAT6, CFP10, MPB83) and the TB Stat-Pak assay were used as surrogate serological markers for TB infection in elephants. In view of the low number of animals of which the infected status could be confirmed (4 out of 708) Latent Class Analyses of TB serology test outcomes was used to predict the putative TB status of each of 708 elephants as positive (17.3%), inconclusive (48.7%), or negative (34%) when assessed on a population basis. Correlation between test performance of the individual assays was high between the ELISAs, but low with that of the TB Stat-Pak assay. Risk factors, assessed based on cut off values for each of the ELISAs determined by ROC analysis, included sex, BCS, age, working time, feed type, management system, camp size and region. Old age elephants were more likely to show a positive TB serology test outcome, than younger ones. Elephants working 7 h per day and the ones in good condition BCS (7–11) were less likely to be positive in TB serology testing. In addition, fewer animals in the large camp size (31–50 elephants) were found to be positive in ELISA tests, compared to elephants in the other camp sizes. In this study, the North region had the lowest percentages of elephants with positive TB test outcome, the West region and to a lesser extend the other regions showed clearly higher percentages of positive animals. Even though assays used in the present study have not been validated yet, results obtained showed promise as diagnostic or screening tests. For the diagnosis of animals suspected to be infected, the ELISA tests, once further optimized for the individual antigens, can be used in parallel. For screening of complete camps for presence or absence of infection, a single optimized ELISA test can be utilized.Faculty of Veterinary Medicine, Chiangmai Universityhttps://www.frontiersin.org/journals/veterinary-science#am2022Veterinary Tropical Disease

Mycobacterium tuberculosis Infection of Domesticated Asian Elephants, Thailand

Four Asian elephants were confirmed to be infected with Mycobacterium tuberculosis by bacterial culture, other diagnostic procedures, and sequencing of 16S–23S rDNA internal transcribed spacer region, 16S rRNA, and gyrase B gene sequences. Genotyping showed that the infectious agents originated from 4 sources in Thailand. To identify infections, a combination of diagnostic assays is essential

Detection of virus-neutralising antibodies and associated factors against rabies in the vaccinated household dogs of Kathmandu Valley, Nepal.

BackgroundRabies is a vaccine-preventable neglected tropical viral zoonosis. It occurs worldwide, creating a very heavy burden in many developing countries, including Nepal. Dogs are the principle vector for the transmission of this disease in urban areas. Vaccination is the most important preventive measure in areas where dogs are the principle source of infection. This study was conducted with the aim of detecting virus-neutralising antibodies and associated factors against rabies in vaccinated household dogs of Kathmandu valley.MethodsBlood samples were collected from 110 vaccinated pet dogs in Kathmandu, Bhaktapur, and Lalitpur districts of Nepal. The samples were taken to the laboratory of the National Zoonosis and Food Hygiene Research Center where serum was separated. An indirect immune-enzymatic assay (PlateliaTM Rabies II kit ad usum Veterinarium, Biorad, China) was used for the detection of rabies virus anti-glycoprotein antibodies in the dog serum samples following the manufacturer's recommendations and instructions. Optical density values for unknown samples were compared with the positive sera titers in quantification tests obtained after a direct reading on the standard curve. Results were expressed as equivalent units per ml (EU/ml).FindingsOf the total samples, 89.09% exceeded the required seroconversion level (≥ 0.5 EU/ml); another 9.09% did not reach the seroconversion level (0.125-0.5 EU/ml); and 1.81% had undetectable seroconversion levels (InterpretationVaccination is the most effective measure for prevention and control of rabies. The locally manufactured brand of vaccine, which is available in Nepal, is potent enough to generate a sufficient amount of protective antibodies, equal to international brands