Bacteriology
Detection of Mycoplasma bovis with an
improved pcr assay. M. Tenk, Á. Bálint, L. Stipkovits, Judit Bíró and L. Dencső
427
Differentiation
of Mycoplasma gallisepticum strains using molecular methods. Judit Bíró, Noémi
Erdei, Ibolya Székely and L. Stipkovits 437
Cloning
and characterisation of the ahpa gene of Pasteurella multocida serogroup b:2
(strain p52): Short
communication. S. Sudha Rani, V. K. Chaturvedi, P. K. Gupta, S. Joseph, B. C. Nair and K. D. Pandey
449
Biochemistry and Physiology
The chicken pituitary-specific transcription factor pit-1 is involved in the hypothalamic regulation
of pituitary hormones. P. Van As, K. Janssens, K. Pals, B. De Groef,
O. M. Onagbesan, V. Bruggeman, V. M. Darras, C. Denef and E. Decuypere 455
Clinical veterinary medicine
Recent developments in canine atopic dermatitis: A review. Noémi Tarpataki 473
Parasitology
Seroprevalence of neosporosis in beef and dairy cattle breeds
in Northeast Hungary. S. Hornok, Renate Edelhofer and I.
Hajtós 485
Babesia divergens becoming
extinct in cattle of Northeast
Hungary:
new data on the past and present situation. S. Hornok, Renate
Edelhofer, I.
Szotáczky and I. Hajtós
493
Pathology
Epidemiological and pathological study on the causes of
abortion in sheep and goats in Hungary (1998–2005). L. Szeredi, Sz. Jánosi, M. Tenk, L. Tekes, M.
Bozsó, Z. Deim and T. Molnár
503
Detection
of Helicobacter
candidatus suis
by pcr in oesophagogastric ulcers of swine
in Italy.
Simonetta
Appino, F. Guarda, Paola Pregel, S. Amedeo, M. A. Cutufia, Giuseppina Bellonio
and A. Ponzetto
517
Virology
Application
of real-time rt-pcr utilising
lux
(light upon extension) fluorogenic primer for the rapid detection of avian influenza
viruses. I. Kiss, P. Germán, L. Sámi, Márta Antal, T. Farkas, G. Kardos, S.
Kecskeméti, Á. Dán and S.
Belák
525
DETECTION OF MYCOPLASMA
BOVIS WITH AN IMPROVED PCR ASSAY
M. Tenk1, Á. Bálint1, L. Stipkovits2, Judit Bíró2 and L. Dencső1
1Central Veterinary
Institute, H-1149 Budapest, Tábornok
u. 2, Hungary; 2Veterinary
Medical Research Institute of the Hungarian Academy of Sciences, Budapest, Hungary
(Received 6 December 2005; accepted 13
February 2006)
A Mycoplasma
bovis species-specific PCR assay has been developed with improvement of a previously
described method (Ghadersohi et al., 1997). This test and its semi-nested version (Hayman
and Hirst, 2003) did not function at all in our hands. A new reverse primer (Mbr2) was
designed using previously published sequence data. For testing specificity, DNA was
extracted from the most frequently occurring mycoplasma species and bacteria of bovine
origin. The new PCR detected only Mycoplasma bovis.
Moreover, no cross-reaction was observed with the genetically closest relative species, M. agalactiae. The target organism could be
detected in a dose as low as 150 CFU ml–1 in broth cultures using
ethidium-bromide-stained agarose gels.
Key
words: Mycoplasma bovis, detection, PCR,
diagnostic assay
DIFFERENTIATION OF MYCOPLASMA
GALLISEPTICUM STRAINS USING MOLECULAR METHODS
Judit Bíró, Noémi Erdei, Ibolya Székely and L. Stipkovits
Veterinary Medical Research
Institute of the Hungarian Academy of Sciences, H-1581 Budapest, P.O. Box 18, Hungary
(Received 13 January 2006;
accepted 13 February 2006)
Increasing use of Mycoplasma gallisepticum (MG) live vaccines has
led to a need for the differentiation of MG strains. The MG strains MK-7, MS-16, S6, FS-9
and R strains and the MG live vaccine strain F were compared by random amplification of
polymorphic DNA (RAPD) in this study. Using RAPD, different patterns were found among the
MG strains. In addition to this, we examined the differentiating potential of polymerase
chain reaction-restriction fragment length polymorphism (PCR-RFLP) primers targeted at the
crmA,
crmB, crmC, gapA, mgc2
and pvpA genes encoding cytadherence-related
surface proteins. These proteins may take part in the pathogenesis of MG-induced disease.
Differentiation of strain F is based on the identification of restriction enzyme sites in
the PCR amplicons. Using HphI enzyme, crmC PCR amplicons produced different RFLP
patterns. Digestion of amplicons of gapA-specific
PCR with MboI enzyme also produced distinct
patterns. Differences were observed among strains R and F by digestion of mgc2 PCR amplicons with HaeIII and VspI
enzymes and digestion of pvpA PCR amplicons with
AccI, PvuII
and ScrFI endonucleases. This method can be
used for the rapid differentiation of vaccine strain from wild strains. Differentiation of
MG strains is a great advantage for diagnosticians or practitioners and it is useful for
epidemiological studies.
Key
words: Mycoplasma gallisepticum, vaccine,
RAPD PCR, PCR-RFLP
CLONING AND CHARACTERISATION OF THE ahpA GENE OF PASTEURELLA MULTOCIDA SEROGROUP B:2 (STRAIN P52):
SHORT COMMUNICATION
S. Sudha Rani, V. K. Chaturvedi, P. K. Gupta, S. Joseph, B. C. Nair
and K. D. Pandey
N. P. R. E., Indian Veterinary
Research Institute, Izatnagar, Bareilly,
243122 UP, India
(Received 19 March 2006; accepted 24
May 2006)
Pasteurella multocida B:2 is responsible for
haemorrhagic septicaemia in cattle and buffaloes, causing severe economic losses in the
developing countries. In the present study, the ahpA
gene of P. multocida B:2 (P52) was
cloned, sequenced and compared with the previously reported ahpA gene sequence in P. multocida A:1, which is responsible for its
haemolytic phenotype. E. coli DH5? cells were
further transformed with recombinant plasmid carrying the ahpA gene from P. multocida B:2 (P52) but SDS-PAGE
analysis failed to show the expression of haemolysin protein. Slight haemolysis was albeit
observed in horse blood agar plates streaked with recombinant E. coli carrying the ahpA gene. Our study indicates that there is 99.6%
similarity and 0.4% divergence between ahpA
gene of P. multocida B:2 (P52) and P. multocida A:1, while membrane topology analysis
has predicted that ahpA is an inner membrane
protein with two strong hydrophobic regions at the N and C terminals. The presence of
significant homology in ahpA sequence in A:1 and
B:2 perhaps suggests a common mechanism of pathogenesis in different species of animals.
Key
words: Pasteurella multocida B:2, ahpA gene, haemolysis, accession no. AY776332
THE CHICKEN
PITUITARY-SPECIFIC TRANSCRIPTION FACTOR PIT-1 IS INVOLVED IN THE HYPOTHALAMIC REGULATION
OF PITUITARY HORMONES
P. Van As1,
K. Janssens2, K. Pals2, B. De
Groef3, O. M. Onagbesan1, V. Bruggeman1, V. M. Darras3, C. Denef2 and E. Decuypere1
1Division of
Livestock-Nutrition-Quality, Department of Biosystems, Faculty of Bioscience Engineering,
Katholieke Universiteit Leuven, Kasteelpark Arenberg 30, B-3001, Leuven, Belgium; 2Laboratory
of Comparative Endocrinology, Pharmacology Section, Department of Molecular Cell Biology,
Faculty of Medicine, Katholieke Universiteit Leuven, Leuven, Belgium; 3Animal
Physiology and Neurobiology Section, Biology Department, Faculty of Science, Katholieke
Universiteit Leuven, Leuven, Belgium
(Received 13 January 2006; accepted 13
February 2006)
Pit-1 is a
pituitary-specific POU-domain DNA binding factor, which binds to and trans-activates
promoters of growth hormone- (GH), prolactin- (PRL) and thyroid
stimulating hormone-beta- (TSHß) encoding genes. Thyrotropin-releasing hormone
(TRH) is located in the hypothalamus and stimulates TSH, GH and PRL release from the
pituitary gland. In the present study, we successfully used the cell aggregate culture
system for chicken pituitary cells to study the effect of TRH administration on the
ggPit-1* (chicken Pit-1), GH and TSHß mRNA expression in vitro. In pituitary cell aggregates of
11-day-old male broiler chicks the ggPit-1* mRNA expression was significantly increased
following TRH administration, indicating that the stimulatory effects of TRH on several
pituitary hormones are mediated via its effect on the ggPit-1* gene expression. Therefore,
a semi-quantitative RT-PCR method was used to detect possible changes in GH and TSHß mRNA
levels. TRH affected both the GH and TSHß mRNA levels. The results of this in vitro study reveal that ggPit-1* has a role
in mediating the stimulatory effects of TRH on pituitary hormones like GH and TSHß in the
chicken pituitary.
Key
words: Chicken, pituitary cell aggregates, ggPit-1*, TRH
RECENT DEVELOPMENTS IN
CANINE ATOPIC DERMATITIS: A REVIEW
Noémi Tarpataki
Department and Clinic of Internal
Medicine, Faculty of Veterinary Science, Szent István University, H-1078 Budapest,
István u. 2, Hungary
(Received 20 March 2006; accepted 24
May 2006)
Atopic dermatitis (AD)
is a genetically predisposed inflammatory and pruritic allergic skin disease with
characteristic clinical features. New results on the pathogenesis and therapeutic aspects
are discussed in this review. IgE-mediated hypersensitivity may be involved in the largest
subset of atopic patients, yet there is another subset for which such involvement cannot
be documented. Alterations in epidermal barrier function, priming of cutaneous
antigen-presenting cells with IgE, intrinsic keratinocyte defects, and development of
autoimmunity are also factors that contribute to the primary disease. Polymorphisms in
regions of the genome that are of key importance to the inflammatory response contribute
to the patient’s clinical picture. Secondary infections, especially with Staphylococcus and yeast organisms, strongly modify
or augment the inflammatory response, which changes over time. After the treatment of
secondary infections and skin inflammation the avoidance of causal allergens would prevent
relapse. Another causative therapy is the variously effective allergen-specific
immunotherapy. The newest treatments for canine AD (cyclosporin A and tacrolimus) are
highly effective at suppressing the allergic response and comparable to treatment with
glucocorticoids. Canine AD presents a substantial diagnostic and therapeutic challenge
over a patient’s lifetime, and no single treatment is universally effective.
Key
words: Dog, atopic dermatitis, hypersensitivity, allergy
SEROPREVALENCE OF NEOSPOROSIS IN BEEF AND DAIRY CATTLE BREEDS
IN NORTHEAST HUNGARY
S. Hornok1,
Renate Edelhofer2 and I. Hajtós3
1Department of
Parasitology and Zoology, Faculty of Veterinary Science, Szent István University, H-1078
Budapest, István u. 2, Hungary; 2Department of Pathobiology, Institute of
Parasitology and Zoology, University of Veterinary Medicine, Vienna, Austria; 3Veterinary
Station of Borsod-Abaúj-Zemplén County, Miskolc, Hungary
(Received 2 January 2006; accepted 24
May 2006)
In order to assess the
seroprevalence of bovine neosporosis with indirect fluorescent antibody test (IFAT), blood
samples were collected randomly from 1063 beef and dairy cattle belonging to 12 different
breeds in Northeast Hungary.
Antibodies to Neospora caninum were detected in
27 (2.5%) of the animals, kept on 19 of the 42 settlements included in this survey. Since
samples were collected on 50 farms, herd prevalence amounted to 38%. The percentage of
cattle with seroconversion increased with age, suggesting a postnatal source of infection.
The highest rate of positivity was detected in Aberdeen Angus (3.3%) and Holstein-Friesian
cows (3.2%), and the lowest in Limousine (0.9%), but no breed predisposition was
statistically substantiated. Neosporosis was more prevalent in dairy (3.4%) than in beef
(1.9%) cattle, although the difference was not significant. Only three out of the
seropositive cows, all of them Holstein-Friesians, had a history of abortion.
Key
words: Neospora caninum, cattle,
seroprevalence, breed, dairy, beef
BABESIA DIVERGENS
BECOMING EXTINCT IN CATTLE OF NORTHEAST
HUNGARY: NEW DATA ON THE PAST
AND PRESENT SITUATION
S. Hornok1,
Renate Edelhofer2, I. Szotáczky3 and I. Hajtós3
1Department of
Parasitology and Zoology, Faculty of Veterinary Science, Szent István University, H-1078
Budapest, István u. 2, Hungary; 2Department of Pathobiology, Institute of
Parasitology and Zoology, University of Veterinary Medicine, Vienna, Austria; 3Veterinary
Station of Borsod-Abaúj-Zemplén County, Miskolc, Hungary
(Received 13 January 2006; accepted 24
May 2006)
Previously unpublished
data from 1958 to 1967 attest the occurrence of Babesia
divergens in cattle in several endemic foci of Northeast
Hungary. During that period the number of clinical cases showed
fluctuation with intervals of 4–5 years and monophasic seasonality (peaking in
June). In order to assess the current status of bovine babesiosis in that region, blood
samples were collected from 654 cattle on 44 farms of 36 settlements in or near the
endemic area during 2005, and serum levels of IgG antibodies to B. divergens were measured by indirect fluorescent
antibody test (IFAT). Only 2 samples (0.3%) showed positivity. In one village clinical
babesiosis was observed over the past few years. Animals brought into the endemic area
during the spring developed haemoglobinuria in the summer of the same year, but those
introduced during the summer or autumn showed clinical signs only after two years. Sampled
animals born and raised locally had neither haemoglobinuria nor seroconversion. Reduction
in the number of cases during the past decades may have been influenced by the
availability of hosts (i.e. decrease of cattle breeding) and the activity of vectors
associated with climate-related changes (e.g. increase of annual sunlight hours in the
endemic area). This is the first report on the prevalence of antibodies to B. divergens
in cattle in Hungary.
Key
words: Babesia divergens, prevalence,
seasonality, haemoglobinuria, climate changes
EPIDEMIOLOGICAL AND PATHOLOGICAL STUDY ON THE CAUSES OF
ABORTION IN SHEEP AND GOATS IN HUNGARY (1998–2005)
L. Szeredi, Sz. Jánosi, M. Tenk, L. Tekes, M. Bozsó, Z. Deim and T. Molnár
Central Veterinary Institute,
H-1149 Budapest, Tábornok u. 2, Hungary
(Received 2 January 2006; accepted 24
May 2006)
The objective of the
investigations was to study the causes of abortion in sheep and goats in Hungary during a 7.5-year period. The authors
investigated 246 cases of ovine and 75 cases of caprine abortions by different diagnostic
methods. An infectious origin was found in 126 cases (51.2%) of ovine and 19 cases (25%)
of caprine abortions. The most important cause of ovine and caprine abortions was Chlamydophila abortus infection with a prevalence
of 46% and 17%, respectively. Other infections causing sheep and goat abortions were
present only in 5.2% and 8% of the cases, respectively. The results obtained by different
diagnostic methods are discussed.
Key
words: Sheep, goat, infectious abortion,
pathology, immunohistochemistry
Corresponding author: Levente Szeredi; E-mail:
szeredil@oai.hu; Phone: 0036 (1) 460-6305; Fax: 0036 (1) 222-6071
DETECTION OF HELICOBACTER CANDIDATUS SUIS BY PCR IN
OESOPHAGOGASTRIC ULCERS OF SWINE IN ITALY
Simonetta Appino1,2, F. Guarda1, Paola Pregel1, S. Amedeo1, M. A. Cutufia3, Giuseppina Bellonio1 and A. Ponzetto4
1Dipartimento
di Patologia Animale, Universita degli Studi di Torino, via L. Da Vinci 44, 10095 Grugliasco (Torino), Italy; 2Istituto di Patologia
Generale, Anatomia Patologica e Clinica Ostetrico-Chirurgica Veterinaria, Universita degli
Studi di Sassari, Sassari, Italy; 3Dipartimento di Genetica, Biologia e
Biochimica, Universita degli Studi di Torino, Torino, Italy; 4Dipartimento di
Gastro-Epatologia, Molinette Hospital, Torino, Italy
(Received 30 January 2006; accepted 24
May 2006)
The aim of this study
was to evaluate by PCR the presence of Helicobacter
spp. in gastric mucus from the fundic region of the stomach and to investigate its role in
oesophagogastric ulcers in swine bred and regularly slaughtered in Piedmont (Northern Italy). Stomachs from 595 regularly slaughtered swine were subjected to gross
pathological examination in order to evaluate the presence of gastric ulcers (revealed in
75 cases, 12.6%). Histopathological examination was performed to better characterise
erosions and ulcers. DNA extracted from gastric mucus collected from all the
ulcer-affected and from 25 normal stomachs was submitted to PCR using Helicobacter spp. 16S rRNA gene target primers.
Sixty-three percent (47/75) of the affected stomachs was positive as well as 24% (6/25) of
the non-affected ones. Sequence analysis from 5 positive samples showed 99% homology with Helicobacter candidatus suis 16S ribosomal RNA
gene.
Key
words: Swine, gastric ulcer, Helicobacter,
PCR
APPLICATION OF REAL-TIME RT-PCR UTILISING LUX (LIGHT UPON
EXTENSION) FLUOROGENIC PRIMER FOR THE RAPID DETECTION OF AVIAN INFLUENZA VIRUSES
I. Kiss1,, P. Germán1, L. Sámi1, Márta Antal1, T. Farkas1, G. Kardos1,2, S. Kecskeméti1, Á. Dán3 and S. Belák4
1Central
Veterinary Institute, Institute of Debrecen, H-4031 Debrecen, Bornemissza u. 3–7, Hungary; 2Department of Medical Microbiology, University of
Debrecen, Debrecen, Hungary; 3Central Veterinary Institute, Budapest, Hungary; 4Departments
of Virology, National Veterinary Institute and Swedish University of Agricultural
Sciences, Uppsala, Sweden
(Received 29 May 2006; accepted 20
September 2006)
A real-time RT-PCR assay
utilising light upon extension fluorogenic primer (LUX RT-PCR) was developed for the rapid
and efficient detection of avian influenza viruses (AIV). The assay detected each of the
AIV isolates tested (16/16) and gave negative results with heterologous pathogens (17/17).
The detection limit of the assay proved to be 10–0.5 EID50/0.2 ml
and 101.5 EID50/0.2 ml in allantoic fluid of virus-infected
embryonated chicken eggs and in spiked chicken faeces samples, respectively. Based on its
specificity, sensitivity and relative simplicity, the LUX RT-PCR assay provides a novel,
rapid and cost-effective diagnostic tool for avian influenza surveillance and monitoring
programs.
Key
words: Avian influenza virus, LUX RT-PCR,
rapid detection
