Andrej Bidovec

Transfer of maternal immunoglobulins and antibodies to Mycoplasma gallisepticum and Mycoplasma synoviae to the allantoic and amniotic fluid of chicken embryos

Maternal antibodies can protect avian embryos against vertically transmitted pathogens during embryogenesis and also young birds after hatching. In contrast to the well-known transfer of maternal immunoglobulin (Ig) G (also termed IgY) from the yolk to embryonic blood, information about the transfer of IgA, IgG and IgM from the egg albumen to the extra-embryonic fluids is very limited. In our study, IgA, IgG and IgM to Mycoplasma gallisepticum and Mycoplasma synoviae were detected in oviduct washings of naturally infected hens and in the corresponding egg albumen samples. In their progeny embryos, IgA, IgG and IgM antibodies to these Mycoplasma species were detected in the allantoic fluid (ALF) and amniotic fluid (AMF) on day 14 of embryonic development (ED). Examination of embryos from chickens immunized with antigens of M. synoviae revealed that the appearance of IgA and IgG and of antibodies to M. synoviae in ALF could vary even among embryos of the same dam. However, IgA, IgG and IgM were detected as early as day 7 of ED in ALF and AMF in certain embryos from hens infected with M. synoviae. In five groups of embryos examined on day 7, IgG to M. synoviae was found in 51% of ALF and 33% of AMF samples. M. synoviae was isolated from 2.3% of ALF samples. IgA to M. synoviae appeared in ALF and AMF on day 12 of ED, and could be found in the majority of AMF samples examined from day 14 onwards. IgM to M. synoviae appeared in AMF on day 13 and in ALF on day 14, but was detected in those fluids less frequently than IgA or IgG. Transfert des immunoglobulines maternelles et des anticorps de Mycoplasma gallisepticum et de Mycoplasma synoviae dans les liquides allantoïque et amniotique des embryons de poulet Les anticorps maternels peuvent protéger les embryons de poulet, contre les agents pathogènes transmis verticalement durant lembryogenèse, et également les jeunes sujets après léclosion. Le transfert des immunoglobulines G maternelles (IgG, également nommées IgY) du vitellus au sang embryonnaire est bien documenté, alors que le transfert des IgA, IgG et IgM de lalbumen aux liquides extra embryonnaires lest très peu. Dans notre étude, les IgA, IgG et IgM vis-à-vis de Mycoplasma gallisepticum et Mycoplasma synoviae ont été détectées dans les lavages doviducte des poules naturellement infectées et dans les échantillons dalbumen des œufs des mêmes sujets. A partir des embryons correspondant aux mêmes sujets, les anticorps IgA, IgG et IgM spécifiques de ces mycoplasmes ont été détectés dans le liquide allantoïdien (ALF) et dans le liquide amniotique (AMF), le 14ème jour du développement embryonnaire (ED). Lexamen des embryons issus de poules immunisées avec les antigènes de M. synoviae a révélé que lapparition dIgA et dIgG et danticorps de M. synoviae dans lALF pouvait varier parmi les œufs dune même poule. Cependant, les IgA, IgG et IgM ont été détectées dès le 7ème jour de lED dans lALF et lAMF de quelques embryons de poules infectées par M. synoviae. Dans cinq groupes dembryons examinés le 7ème jour, les IgG spécifiques de M. synoviae ont été détectées dans 51% des échantillons dALF et 33% de ceux dAMF. M. synoviae a été isolé à partir de 2,3% des échantillons dALF. Les IgA spécifiques de M. synoviae sont apparues dans les ALF et AMF le 12ème jour de lED et ont pu être détectées dans la majorité des échantillons dAMF examinés à partir du 14ème jour. Les IgM spécifiques de M. synoviae sont apparus dans les AMF le 13ème jour et dans les ALF le 14ème jour, mais ont été détectées dans les liquides moins fréquemment que les IgA ou IgG. Übertragung maternaler Immunglobuline und Antikörper gegen Mycoplasma gallisepticum und Mycoplasma synoviae in die Allantois- und Amnionflüssigkeit des Hühnerembryos Maternale Antikörper können Vogelembryonen während der Embryogenese gegen vertikal übertragene Pathogene und auch Jungvögel nach dem Schlupf gegen Krankheitserreger schützen. Im Gegensatz zu der bekannten Übertragung des maternalen Immunglobulins G (IgG, auch als IgY bezeichnet) vom Dotter in das embryonale Blut ist das Wissen über den Transfer von IgA, IgG und IgM vom Eialbumin in die extraembryonalen Flüssigkeiten sehr begrenzt. In unserer Studie wurden IgA, IgG und IgM gegen Mycoplasma gallisepticum und Mycoplasma synoviae in Eileiterspülflüssigkeiten von natürlich infizierten Hennen sowie in den korrespondierenden Eialbuminen nachgewiesen. In den embryonalen Nachkommen dieser Hennen wurden am 14. Tag der embryonalen Entwicklung (ED) IgA-, IgG- und IgM-Antikörper gegen diese Mycoplasma-Spezies in der Allantoisflüssigkeit (ALF) und der Amnionflüssigkeit (AMF) festgestellt. Die Untersuchung von Embryonen von Hennen, die mit M. synoviae-Antigen immunisiert worden waren, ließen erkennen, dass das Auftreten von IgA und IgG sowie von Antikörpern gegen M. synoviae in der ALF, sogar zwischen Embryonen von derselben Henne, variieren konnte. IgA-, IgG- und IgM konnte jedoch in bestimmten Embryonen von M. synoviae infizierten Hennen bereits am 7. Tag der ED in der ALF und AMF entdeckt werden. In fünf am 7. Tag untersuchten Gruppen von Embryonen wurden IgG gegen M. synoviae in 51 % der ALF- und 33 % der AMF-Proben gefunden. M. synoviae wurde aus 2,3 % der ALF-Proben isoliert. IgA gegen M. synoviae erschien in der ALF und AMF am 12. Tag der ED und konnte ab dem 14. Tag in der Mehrzahl der untersuchten AMF-Proben nachgewiesen werden. IgM gegen M. synoviae trat in der AMF ab dem 13. und in der ALF ab dem 14. Tag der ED auf, wurde aber in diesen Flüssigkeiten weniger häufig gefunden als IgA und IgG. Transferencia de inmunoglobulinas maternales y anticuerpos frente a Mycoplasma gallisepticum y Mycoplasma synoviae a los líquidos alantoideos y amnióticos de embriones de pollo Los anticuerpos maternales protegen a los embriones de ave de los patógenos que se transmiten vía vertical durante la embriogénesis y también a los pollitos tras el nacimiento. Aunque la transferencia de inmunoglobulinas maternales (IgG, también llamadas IgY) de la yema a la sangre del embrión se conoce bien, la información existente sobre la transferencia de IgA, IgG e IgM desde el albumen del huevo hasta el líquido extraembrionario es limitada. En nuestro estudio, se detectaron las IgA, IgG e IgM frente a Mycoplasma gallisepticum y Mycoplasma synoviae en lavados de oviducto de gallinas infectadas de forma natural y en las correspondientes muestras de albumen del huevo. En los embriones de la progenie de estas gallinas se detectaron anticuerpos IgA, IgG e IgM frente a estas especies de Mycoplasma en el líquido alantoideo (ALF) en líquido amniótico (AMF) en el día 14 del desarrollo embrionario (ED). El examen de los embriones de pollos inmunizados con antígenos de M. synoviae revelaron que la aparición de anticuerpos IgA e IgG frente a M. synoviae en ALF puede variar incluso entre embriones de las misma reproductora. Aún así, las IgA, IgG e IgM se detectaron tan pronto como el día 7 de ED en ALF y AMF en ciertos embriones de gallinas infectadas con M. synoviae. En cinco grupos de embriones examinados el día 7, se detectaron IgG frente a M. synoviae en el 51% de ALF y en el 33% de las muestras de AMF. M. synoviae fue aislado de 2.3% de las muestras de ALF. Las IgA frente a M. synoviae aparecieron en ALF y AMF el día 12 de ED y pudieron ser encontradas en la mayoría de las muestras a partir del día 14. Las IgM frente a M. synoviae aparecieron en el AMF el día 13 y en ALF el día 14, pero fue detectado en estos fluidos menos frecuentemente que las IgA o IgG.

Fasciola hepatica: effects on blood constituents and liver minerals in fallow deer (Dama dama)

Haematological and serum biochemical constituents as well as the liver mineral concentrations can be affected by Fasciola hepatica. The present study compares the blood and liver constituents in 19 fallow deer with fascioliosis and in 43 without it. Blood samples, gastro-intestinal tract, lungs and liver were taken from 62 male and female fallow deer that were shot, aged 6 months and over, in enclosures, between 1999 and 2001 in Slovenia. Samples were analysed and the means of various parameters of blood and liver were determined. Significant differences in blood were found in the mean cell volume, total serum proteins, sodium, and calcium; and in liver and calcium and copper were higher in animals without fascioliosis. Mean cell haemoglobin concentration, eosinophils, basophils, lymphocytes, monocytes, alanine aminotransferase, urea, glucose and copper in blood were significantly higher in animals with fascioliosis. It has been concluded that some of the values in fallow deer like in other domestic ruminants are the sensitive indicators of liver cell damage in fascioliosis.

Presence of Antibodies Against Aujeszky's Disease Virus in Wild Boar (Sus scrofa) in Slovenia

Serum samples from 427 hunter-killed wild boar (Sus scrofa) from Slovenia were tested for antibodies to Aujeszkys disease virus (ADV). Samples were collected throughout Slovenia and corresponded to 6.2% of the total harvest. Antibodies against ADV were detected in 111 sera (26%) using a commercial enzyme-linked immunosorbent assay (ELISA). Antibody prevalence increased significantly with age. This report describes the first evidence of ADV infection in wild boar populations in Slovenia.

Echinococcus multilocularis in the red fox (Vulpes vulpes) in Slovenia

Using the parasitological washing out method, we examined the intestines of 428 red foxes (Vulpes vulpes) for the presence of Echinococcus multilocularis (Leuckart 1863) and found that the overall prevalence was 2.6% (confidence interval 95% 1.3–4.5%). This is the first extended research reporting on the presence of E. multilocularis in the Slovenian fox population.

Control of Rabies in Slovenia

Red foxes (Vulpes vulpes) are the main reservoir of rabies in Slovenia, whereas cases of rabies in other wildlife species occur sporadically. In 1995, a program of oral vaccination of wildlife in Slovenia was initiated; baits with oral vaccine were distributed by air at a density of 20 baits/km2. During 1995, when the oral vaccination program was started, 1,089 cases of rabies (including both wild and domestic animals) were reported. Five years later (1999), only six positive animals were detected among 1,195 tested (0.5%). Despite an increase in bait density (25 baits/km2) during the years 2000 and 2001, reported rabies cases increased to 115 and 135, respectively. In 2003, following initiation of a new bait-dropping strategy, which incorporated perpendicular rather than parallel flight lines, the number of rabies cases decreased to eight.

Leptospira antibodies in wild boars (Sus scrofa) in Slovenia

Serum samples collected from 437 shot wild boars (Sus scrofa) were tested for the presence of antibodies against Leptospira interrogans sensu lato in wild boar in Slovenia. Assessment of leptospira-specific antibodies was performed by microscopic agglutination test. Antibodies against at least one of the pathogenic serovars were detected in 200 (45.8%) sera. From 200 positive samples, 100 samples (50%) had positive titre against a single serovar, while 100 (50%) samples had positive titres against two or more serovars. The most frequently detected antibodies were those against serovar Tarassovi. This investigation confirmed the presence of different pathogenic serovars in wild boar across Slovenia. It can be concluded that wild boars are natural reservoirs of at least some of the leptospiral serovars that represent a potential source of leptospirosis for other wild and domestic animals, as well as for humans.

Immunohistochemical Study of Normal and Mange (S. scabiei var. rupicaprae) Infested Chamois (Rupicapra rupicapra L.) Skin

The immunohistochemical study of chamois (Rupicapra rupicapra L.) skin showed that a limited number of available monoclonal and polyclonal antibodies expressed reactivity with skin cell components. These included cytokeratins, vimentin, desmin, neuron‐specific enolase and S‐100 protein with almost the same distribution pattern as already described in the skin of humans and animals. Antibodies used for labelling skin‐associated lymphoid tissues and other cells with the immuno‐logic function in human skin failed to demonstrate these cells in the chamois skin with the exception of LCA and OKT6 antibodies. Epidermal Langerhans cells were reliably demonstrated only by the enzyme histochemical method for adenosine triphosphatase, while the majority of mononuclear cells in dermal infiltrates showed a strong immunoreaction with OKT6 antibody. The histologic and histochemical analysis showed that the dermal infiltrations in infested skin consisted of macro‐phages, lymphocytes, granulocytes, mastocytes and fibroblasts. The chamois skin affected with sarcoptes mange showed a significant loss of cytokeratins in the epidermis and its derivatives. Particular keratinocytes showing nonspecific staining with several antibodies were also described and discussed in this paper.

Biochemical parameters in blood of farmed fallow deer (Dama dama)

SummaryWe analyzed blood samples from 62 shot fallow deer and determined various biochemical parameters. The blood samples were taken during winter from individuals living in enclosures in several regions of Slovenia during winter. The results have been analyzed according to sex and age of the animals. The following parameters were determined: aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), gamma glutamyltransferase (GGT), glutamate dehydrogenase (GLDH), urea, creatinine, total protein, albumin, glucose, iron (Fe), sodium (Na), potassium (K), chloride (Cl), calcium (Ca), inorganic phosphorus (aP), magnesium (Mg) and copper (Cu). Minor significant differences in enzyme activities and electrolyte concentrations were found between sexes and age groups.ZusammenfassungWir bestimmten verschiedene biochemische Kennwerte in Blutproben von 62 geschossenen Damhirschen. Die Blutproben wurden im Winter genommen und stammten von Gehegetieren aus verschiedenen Regionen Sloweniens. Die Ergebnisse wurden im Hinblick auf Geschlechts- und Altersunterschiede analysiert. Es wurden folgende Parameter untersucht: Aspartat Aminotransferase (AST), Alanin Aminotransferase (ALT), Lactat Dehydrogenase (LDH), Gamma Glutamyl Transferase (GGT), Glutamat Dehydrogenase (GLDH), Harnstoff, Kreatinin, Gesamtprotein, Albumin, Glucose, Eisen, Natrium, Kalium, Chlorid, Kalzium, anorganischer Phosphor, Magnesium und Kupfer. Es wurden geringe signifikante Unterschiede in Enzymaktivität und Elektrolytkonzentrationen zwischen den Geschlechtern und Altersgruppen ermittelt.RésuméDes échantillons de sang obtenus auprès de 62 daims tirés en Slovénie ont été analysés et les moyennes de différents paramètres biochimiques ont été déterminées. Les échantillons provenaient danimaux tirés en hiver dans des enclos situés dans différentes régions de la Slovénie. Les résultats ont été analysés en fonction du sexe et de lage. Différents groups ont été comparés sur base des caractéristiques prises en considération. Les paramètres suivants ont été déterminés: aspartate aminotransférase (AST), alanine aminotransférase (ALT), lactate déshydrogénase (GLDH), résine, créatinine, protéine totale, albumine, glucose, Fer (Fe), Sodium (Na), Potassium (K), Chlore (Cl), Calcium (Ca), Phosphore anorganique (aP), Magnésium (Mg) et Cuivre (Cu). Des différences significatives mineures ont été trouvées en ce qui concerne les électrolytes et lactivité des enzymes en classant les échantillons par sexe et par âge.

Stichorchis subtriquetrus in two beavers (Castor fiber) from Slovenia

SummaryIntestines of two beavers (Castor fiber), one killed by a car accident and one drowned were examined for parasites. Examination revealed the presence of 1 and 14 adult trematodes, Stichorchis subtriquetrus in the caecum and colon, respectively. The flotation method revealed no ova from faecal material. The European beaver was present in the territory of Slovenia until the beginning of the 18th century. Recently, in 1998, the species reappeared in southeast of Slovenia as a result of the reintroduction of European beaver in Croatia. This is the first report of veterinary examination of beaver and the first evidence of S. subtriquetrus in Slovenia.

Blood and urine values of free-living European wildcats in Slovenia

Hematological, serum biochemistry, and urinalysis values were determined for nine (two females and seven males) adult, free-living European wildcats (Felis silvestris) in the Kocevje Forests of the southern Slovenia. Samples were collected from August 1999 to March 2001. Cats were anesthetized with ketamine and medetomidine. Blood samples were taken by jugular venipuncture and urine samples by bladder puncture. A control group of domestic cats (F. silvestris catus) was assembled to determine if differences exist among blood and urine values between free-living European wildcats and domestic cats. Hematological, biochemical, and urine parameters were similar to those of the control group. Values of glucose, blood urea nitrogen, albumin, mean corpuscular value, basophile count, and alanine aminotransferase were significantly higher than values of the control group. All urine samples contained white blood cells and proteins, and seven of them contained red blood cells.