Investigating the Prevalence of Paratuberculosis in Hungarian Large-Scale Dairy Herds and the Success of Control Measures over Four Years.

Paratuberculosis (PTB) is a severe, slow-developing, untreatable disease of ruminants. Worldwide, the disease affects more than 50% of herds in the dairy industry, and causes substantial economic losses for dairy producers. Diagnostic tests show limited sensitivity, especially in the early stages of the disease. Our study aimed to investigate the seroprevalence of Mycobacterium avium ssp. paratuberculosis (MAP) in large-scale dairy herds in Hungary, in association with the self-reported presence or absence of screening and intervention measures against MAP transmission. We processed data from 42 large-scale Holstein Friesian farms in Hungary between 1 January 2018 and 31 December 2021. An average of 32,009 (min.: 31,702; max.: 32,207) animals were blood sampled yearly (127,372 in total during the four years), corresponding to 15% of the Hungarian dairy cattle population. All female cattle older than 2 years were blood sampled on the farms enroled in the study. The samples were tested using a commercial ELISA (IDEXX paratuberculosis screening Ab test). Farm managers were interviewed about their on-farm diagnostic and intervention approaches using a uniform questionnaire, including questions on the level of awareness, frequency of ELISA and PCR testing, and their strategies for culling adult animals and reducing transmission to newborn calves. By comparing the annual rate of change in seroprevalence and the amount of change observed during the four-year period, we concluded that test-and-cull strategies implemented in parallel with newborn calf management that aimed at preventing MAP transmission were superior to test-and-cull strategies alone; moreover, fortifying culling decision making via additional ELISA and PCR tests is superior to using a single ELISA result. For farms that carried out a complex program with both "test-and-cull" and proper newborn calf management, there was a proportional reduction in apparent seroprevalence at an average of 22.8% per year. Fifteen of the sampled farms had no measures in place to control paratuberculosis. On these farms, the seroprevalence increased by 12.1% per year on average.

New record of metacercariae of the North American Posthodiplostomum centrarchi (Digenea, Diplostomidae) in pumpkinseed (Lepomis gibbosus) in Hungary.

Two species of the genus Posthodiplostomum (Digenea: Diplostomatidae) (Posthodiplostomum brevicaudatum Nordmann, 1832 and Posthodiplostomum cuticola Nordmann, 1832) are known as parasites of Hungarian native fishes. Metacercariae of P. cuticola are widespread in Europe and cause black spot disease. Several species of Posthodiplostomum were described also from North America but none of them has been isolated in Hungary up to now. Posthodiplostomum centrarchi Hoffman, 1958 has been detected recently in pumpkinseeds (Lepomis gibbosus L., 1758) in several European countries. Posthodiplostomum centrarchi was isolated for the first time in Hungary from pumpkinseeds caught in the Maconka water reservoir in 2015. Thereafter, several natural waters (e.g. the River Danube, Lake Balaton and the Sió channel) were sampled in order to determine its presence and distribution. Only the native species P. cuticola was detected in Lake Balaton on cyprinids but a relatively high infection rate of P. centrarchi was observed in the Sió channel close to the lake. Pathological changes were absent, and metacercariae were mostly attached to the surface of the liver, kidney and heart. The phylogenetic analysis of the ITS and COI sequences of P. centrarchi and P. cuticola clustered into two distinct branches, which was in agreement with the morphological results.

Description of two new species of Myxobolus Bütschli, 1892, M. peleci n. sp. and M. cultrati n. sp., detected during an intensive mortality of the sichel, Pelecus cultratus (L.) (Cyprinidae), in Lake Balaton, Hungary.

In the summer of 2014, mass mortality of the sichel, Pelecus cultratus (L.), was observed in Lake Balaton, Hungary. Parasitological examination conducted in the framework of a complete diagnostic survey revealed myxozoan infections. Two species of Myxobolus Bütschli, 1892 were found, one in the gill lamellae and another in the eyes. Following this mass mortality, 113 sichel specimens were examined during a 14-month period. Gill infection with scattered spores in the lamellae was found in 51 fish, while infection in the eyes was recorded in three specimens only. Based upon the morphological and molecular biological data the species from the gills is described here as Myxobolus peleci n. sp. and the species from the eye as M. cultrati n. sp. The 18S rDNA sequences of the two species proved that they differ from all known Myxobolus spp. with sequence data available in the GenBank database. Histological examinations revealed that the spores found in the gill lamellae were derived from plasmodia developing in and around the afferent branchial arteries of the gill arches. No mortality of sichel was recorded in 2015. Infection with these two Myxobolus spp. does not seem to play a role in the mortality of the host fish.

Isolation and characterisation of flavobacteria from wild and cultured freshwater fish species in Hungary.

The objective of this study was to survey the incidence of Flavobacterium columnare in wild and cultured freshwater fish species in Hungary. This bacterium usually causes disease in waters of more than 25 °C temperature. However, with the introduction of intensive fish farming systems, infected fish exposed to stress develop disease signs also at lower temperatures; in addition, the temperature of natural waters rises to the critical level due to global warming. Twenty-five isolates from wild and cultured freshwater fishes were identified as F. columnare by specific PCR, although both the fragment lengths and the results of PCRRFLP genotyping with BsuRI (HaeIII) and RsaI restriction enzymes raised doubts regarding this species classification. Sequencing of the 16S ribosomal RNA gene revealed that 23 isolates belonged to the species F. johnsoniae and two represented Chryseobacterium spp. The isolates were found to have high-level multidrug resistance: all were resistant to ampicillin and polymyxin B, the 23 F. johnsoniae strains to cotrimoxazole, 88% of them to gentamicin, and 72% to chloramphenicol. The majority of the 25 isolates were sensitive to erythromycin (88%), furazolidone (76%), and florfenicol (68%).

The occurrence of metacercariae of Petasiger (Digenea: Echinostomatidae) in an unusual site, within the lateral line scales of cyprinid fishes.

During a regular veterinary inspection of fishes from Lake Balaton, Hungary, echinostomatid metacercariae (Digenea), with collar spines characteristic of species of the genera Petasiger Dietz, 1909 and Paryphostomum Dietz, 1909, were found in the lateral line scales of a roach Rutilus rutilus (Linnaeus), an apparently unique site. In a subsequent examination of 586 fishes from 20 different species, similar infections were found in 11 species. The infection was virtually restricted to the lateral line scales, other scales being infected only incidentally. These encysted metacercariae had 27 collar spines, including eight larger angle spines and 19 smaller dorsal spines arranged in two rows. Two types of metacercarial cyst were found. One type had a cyst diameter of 138-171 µm × 105-120 µm and three central dorsal spines that were larger than the remainder and tended to resemble the angle spines. The second type of metacercarial cyst had a diameter of 128-157 µm × 105-115 µm and all 19 dorsal spines of the metacercaria were of a similar size. ITS sequences of the second type of metacercaria exhibited a 100% similarity to sequences of two adult Petasiger phalacrocoracis (Yamaguti, 1939) specimens collected from the gut of Phalacrocorax carbo (Linnaeus) in Hungary and to P. phalacrocoracis deposited in the GenBank database. Sequences obtained from two metacercariae of the first type showed a 2.8-2.9 % difference from sequences of the second type of metacercaria and from those of adult specimens of P. phalacrocoracis from cormorants. Based on these results, the second type metacercaria is considered to be a larval stage of P. phalacrocoracis, but the identity of the first type is uncertain. The unusual location of these metacercariae in the lateral line scales is discussed in relation to their transmission.

Molecular detection of a putatively novel cyprinid herpesvirus in sichel (Pelecus cultratus) during a mass mortality event in Hungary.

In the early summer of 2014, mass mortality of sichel (Pelecus cultratus) was observed in Lake Balaton, Hungary. Histological examination revealed degenerative changes within the tubular epithelium, mainly in the distal tubules and collecting ducts in the kidneys and multifocal vacuolisation in the brain stem and cerebellum. Routine molecular investigations showed the presence of the DNA of an unknown alloherpesvirus in some specimens. Subsequently, three genes of the putative herpesviral genome (DNA polymerase, terminase, and helicase) were amplified and partially sequenced. A phylogenetic tree reconstruction based on the concatenated sequence of these three conserved genes implied that the virus belongs to the genus Cyprinivirus within the family Alloherpesviridae. The sequences of the sichel herpesvirus differ markedly from those of the cypriniviruses CyHV-1, CyHV-2 and CyHV-3, putatively representing a fifth species in the genus.

Limited macular translocation compared with photodynamic therapy in the management of subfoveal choroidal neovascularization in age-related macular degeneration.

PURPOSE: To compare the visual outcome of macular translocation (MT) versus photodynamic therapy (PDT) for subfoveal predominantly classic neovascularization in age-related macular degeneration (AMD). DESIGN: Nonrandomized clinical trial. METHODS: Retrospective review of 65 consecutive patients with subfoveal neovascularization due to AMD. The follow-up was at least 6 months. Main outcome criteria were final best corrected visual acuity and the gain in visual acuity. RESULTS: A total of 29 eyes were treated with PDT with verteporfin, and 36 underwent MT with chorioscleral infolding. Both groups were similar for age, refraction, and lesion size. The initial visual acuity was lower in the MT group than in the PDT group (20/200 versus 20/100). Mean follow-up was 11 months for the PDT group and 14 months for the MT group. The mean displacement of the fovea after translocation was 1,274 microm (range, 250 to 1,900 microm). Mean number of retreatment by PDT was 2.5. At 1 year, both groups had the same final visual acuity (20/200), but the improvement was more favorable in the MT group (gain of 0.7 line in the MT group versus loss of 3.4 lines in the PDT group, P =.007). One eye in the PDT group (4.3%) had a gain of 3 lines or more versus eight eyes (38%) in the MT group; the lesion size was larger in the PDT group than in MT group (P =.036). CONCLUSION: In this retrospective study, MT seemed to allow a better preservation of visual acuity than PDT in subfoveal neovascularization due to AMD. Further larger and controlled studies are required.

Development of an antibiotic resistance monitoring system in Hungary.

Because of the rapid development and spread of antimicrobial resistance it is important that a system be established to monitor antimicrobial resistance in pathogenic zoonotic and commensal bacteria of animal origin. Susceptibility testing of bacteria from carcasses and different samples of animal origin has been carried out in veterinary institutes for a long time but by an inconsistent methodology. The disc diffusion method proposed by the National Committee for Clinical Laboratory Standards (NCCLS) was introduced in all institutes in 1997. In order to obtain a coherent view of the antimicrobial resistance of bacteria a computer system was consulted, consisting of a central computer to store all data and some local computers attached to it through the network. At these local measuring stations computers are connected to a video camera, which displays the picture of Petri dishes on the monitor, and inhibition zone diameters of bacteria can be drawn with the mouse by the inspector. The software measures the diameters, evaluates whether or not the bacteria are sensitive, and stores the data. The evaluation is based upon the data of the NCCLS. The central computer can be connected to as many local computers with measuring stations as we wish, so it is suitable for an integrated system for monitoring trends in antimicrobial resistance of bacteria from animals, food and humans, facilitating comparison of the occurrence of resistance for each circumstance in the chain. It depends on the examiners which antibiotics they want to examine. Thirty-two different antibiotic panels were compiled, taking into consideration the active ingredients of medicinal products permitted for veterinary use in Hungary, natural resistance and cross-resistance, the mechanism of resistance and the animal species, i.e. which drugs were recommended for treatment in the given animal species, and the recommendations of the OIE Expert Group on Antimicrobial Resistance. The members of the panels can be changed any time, even during the measuring process. In addition to the inhibition zone diameters of bacteria the database also includes information about bacterial and animal species, the age of animals and the sample or organ where the bacteria are from. Since January 2001 the antibiotic susceptibility of E. coli, Salmonella, Campylobacter and Enterococcus strains isolated from the colons of slaughter cows, pigs and broiler chickens has also been examined. Each of the 19 counties of Hungary submits to the laboratory three tied colon samples from a herd of the above-mentioned animals every month.

MRI for the evaluation of scleral invasion and extrascleral extension of uveal melanomas.

AIM: To study the diagnostic value of MRI for the detection of scleral infiltration and extrascleral extension of uveal melanoma. MATERIALS AND METHODS: Twelve consecutive patients with clinically suspected large uveal melanoma were examined by MRI. All of the patients underwent enucleation or exenteration. The clinical and histopathological findings were compared. RESULTS: The clinical diagnosis of uveal malignant melanoma was confirmed by histology in each case. MRI allowed detection of scleral infiltration with a sensitivity of 100% and a specificity of 50%. For extrascleral extension, MRI had a sensitivity of 100% and specificity of 89%. CONCLUSIONS: Using fat suppression technique and enhancement with contrast medium, MRI proved to be a valuable ancillary method for the assessment of scleral infiltration and extrascleral extension.