Health status of free-ranging ring-necked pheasant chicks (Phasianus colchicus) in North-Western Germany.

Being a typical ground-breeding bird of the agricultural landscape in Germany, the pheasant has experienced a strong and persistent population decline with a hitherto unexplained cause. Contributing factors to the ongoing negative trend, such as the effects of pesticides, diseases, predation, increase in traffic and reduced fallow periods, are currently being controversially discussed. In the present study, 62 free-ranging pheasant chicks were caught within a two-year period in three federal states of Germany; Lower Saxony, North Rhine-Westphalia and Schleswig-Holstein. The pheasant chicks were divided into three age groups to detect differences in their development and physical constitution. In addition, pathomorphological, parasitological, virological, bacteriological and toxicological investigations were performed. The younger chicks were emaciated, while the older chicks were of moderate to good nutritional status. However, the latter age group was limited to a maximum of three chicks per hen, while the youngest age class comprised up to ten chicks. The majority of chicks suffered from dermatitis of the periocular and caudal region of the head (57-94%) of unknown origin. In addition, intestinal enteritis (100%), pneumonia (26%), hepatitis (24%), perineuritis (6%), tracheitis (24%), muscle degeneration (1%) and myositis (1%) were found. In 78% of the cases, various Mycoplasma spp. were isolated. Mycoplasma gallisepticum (MG) was not detected using an MG-specific PCR. Parasitic infections included Philopteridae (55%), Coccidia (48%), Heterakis/Ascaridia spp. (8%) and Syngamus trachea (13%). A total of 8% of the chicks were Avian metapneumovirus (AMPV) positive using RT-PCR, 16% positive for infectious bronchitis virus (IBV) using RT-PCR, and 2% positive for haemorrhagic enteritis virus (HEV) using PCR. All samples tested for avian encephalomyelitis virus (AEV), infectious bursal disease virus (IBDV) or infectious laryngotracheitis virus (ILTV) were negative. The pool samples of the ten chicks were negative for all acid, alkaline-free and derivative substances, while two out of three samples tested were positive for the herbicide glyphosate. Pheasant chick deaths may often have been triggered by poor nutritional status, probably in association with inflammatory changes in various tissues and organs as well as bacterial and parasitic pathogens. Theses impacts may have played a major role in the decline in pheasant populations.

Central nervous signs, blindness and cerebral vermicosis in free-ranging peregrine falcons (Falco peregrinus) associated with aberrant larval migrations.

Four free-ranging peregrine falcons (Falco peregrinus) were submitted with a history of unilateral or bilateral blindness and central nervous signs to a veterinary clinic in Germany. There were no indications of trauma or ocular disease. Likewise, other differential diagnoses for CNS signs were ruled out within the diagnostic process. The clinical diagnostic panel in live falcons included general examination, radiography, endoscopy, hematology, ophthalmoscopy and parasitological examination of the feces, blood gas analysis and blood chemistry as well as computed tomography, and magnetic resonance imaging (MRI). A complete pathological and histopathological examination was performed post-mortem. The only common finding in all birds was an infection with the nematode parasite Serratospiculum tendo. The parasite was confirmed morphologically and via PCR. In two falcons intracerebral vermicoses was suspected in MRI and confirmed in subsequent histopathology, but molecular biological identification of the parasite species failed from brain tissue. Until today, S. tendo had been reported to affect the respiratory system, the liver and different parts of the gastrointestinal tract and to cause cachexia, inappetence, regurgitation, dyspnea and general signs of illness such as lethargy, poor plumage, and reduced reproduction. Our findings indicate that aberrant migration should be considered as cause for CNS signs in falcons. As S. tendo might be a possible cause for this, CNS signs might be included in the list of clinical signs of serratospiculiasis in falcons.

Semen analysis and successful artificial insemination in the St. Vincent amazon (Amazona guildingii).

The St. Vincent amazon (Amazona guildingii) is an endemic parrot on the Carribean island St Vincent. Due to poaching, trade, natural events such as hurricanes and habitat loss the species declined severely throughout the 20th century to a total number of 487 adult individuals and is currently regarded as vulnerable by IUCN. Captive breeding is attempted in terms of species conservation, but reproduction rates have been low due to reproductive problems such as mate aggression, asynchronous reproductive activity and infertile eggs. The aims of the present study were; firstly, to evaluate whether semen analysis might help to assess the fertility of males and to detect potential reasons for infertile eggs; secondly, to increase the number of offspring using artificial insemination, and as a future effect, to increase the presence of genetically valuable males in the ex-situ breeding population. For semen collection electric stimulation was used in 15 mature and healthy St. Vincent amazons with a success rate of 89% (202/227 attempts) in 14/15 males. Quality assessment of the semen included the evaluation of volume, pH, color, consistency and contaminations of the ejaculate, as well as estimation of motility, viability, morphology, concentration and total count of spermatozoa. Semen pH ranged from 6.7 to 7.5. Median sperm motility was 50% and median progressively forward motility 40%. Mean sperm concentration (x¯ ± SD) was 21,313.5 ± 22,408.8 spermatozoa/µl and mean sperm viability 66 ± 16%. Semen samples contained on average 20.5% morphologically normal spermatozoa and sperm malformations were detected mainly in the head (x¯ = 47.6%) and the tail regions (x¯ = 27.7%). Interestingly round bodies were detected in the ejaculates with a mean ratio of 0.6 round bodies per sperm. Semen analysis proved to be very useful to identify males with poor semen quality. Artificial insemination was performed 46 times in 9 females with either individual or pooled semen samples and 13 eggs from 7 females were laid afterwards. In 3 eggs, embryonic development was detected and 1 chick hatched successfully. Paternity testing confirmed the fatherhood of a one-winged semen donor male, a bird which was not able to copulate naturally. The results are very promising and underline that assisted reproduction techniques are a suitable tool for species conservation in captive breeding programs for psittacines.

Semen collection, semen analysis and artificial insemination in Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus) as part of a species conservation project.

Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus; hereafter CSTG) have experienced substantial decreases in population numbers and geographic range during the early 20th century, primarily due to habitat loss. The conservation aim of this project was to re-establish a self-sustaining population of CSTG within an unoccupied portion of their historic range in northeastern Nevada via reintroduction from source populations in Idaho, USA. Female nest initiation rates post-translocation due to low fertilization rates are believed to be one limiting factor in the establishment of some translocated CSTG populations. However, studies on semen collection and artificial insemination in this species are absent. Assisted reproduction was evaluated as an additional tool in this species conservation project in order to gain knowledge on the reproductive status of yearling and adult male CSTG, establish orientation values for semen parameters and evaluate artificial insemination procedures on female CSTG. In two consecutive breeding seasons, semen collection was attempted 51 times in 47 males using the established massage method, and a novel electro-stimulation technique. Semen collection was successful in all attempts, even in yearling grouse, which represents a novel confirmation that yearling male CSTG can produce live spermatozoa in their first breeding season. Volume, color, consistency, contamination, pH of semen, and the motility, concentration, viability and morphology of spermatozoa were analyzed. Extracted semen volume ranged between 6 and 74 µl and the mean pH was 6.9 ±â€¯0.5 (x¯ ± SD). Morphology analysis revealed that on average, 42.8% of sperm was morphologically normal, but 34.8% had malformed tails. Additionally, artificial insemination was practiced in 17 females (sham-insemination group; insemination lacking spermatozoa) and performed in 17 females. Intravenous catheters G20 1.0 mm × 32 mm enabled safe intravaginal insemination under visual control. Significant (p ≤ 0.05) differences in semen parameters between adult and yearling birds were detected. It is well established that adult males receive a majority of copulations during lekking, but our novel findings also indicate that they produce significantly more spermatozoa which is of higher quality than yearling males. This finding offers insights into male reproductive biology in a lekking grouse species.

The role of Virus "X" (Tortoise Picornavirus) in kidney disease and shell weakness syndrome in European tortoise species determined by experimental infection.

Tortoise Picornavirus (ToPV) commonly known as Virus "X" was recently discovered in juvenile European tortoises suffering from soft carapace and plastron as well as kidney disease. Therefore, this virus was a potential candidate to be a causative agent for these disease patterns. Spur thighed tortoises (Testudo graeca) seemed to be more susceptible to establish clinical symptoms than other European species like T. hermanni. Thus this trial investigated the role of ToPV in the described syndrome. Two groups of juvenile European tortoises (T. graeca and T.hermanni) each of 10 animals, were cloacally, oronasally and intracoelomically inoculated with an infectious dose (~ 2000 TICD) of a ToPV strain isolated from a diseased T. graeca. A control group of two animals of each species received non-infected cell culture supernatant. The tortoises were examined daily and pharyngeal and cloacal swabs for detection of ToPV-RNA by RT-PCR were taken from each animal every six days for a period of 6 months. At the end of the study the remaining animals were euthanised and dissected. Bacteriological and parasitological tests were performed and organ samples of all tortoises were investigated by RT-PCR for the presence of ToPV and histopathology. Animals that were euthanised at the end of the experiment, were examined for presence of specific anti-ToPV antibodies. Several animals in both inoculated groups showed retarded growth and a light shell weakness, in comparison to the control animals. Three animals were euthanised during the trial, showing reduced weight gain, retarded growth, severe shell weakness and apathy, in parallel to clinical observations in naturally infected animals. In all inoculated animals of both species an intermittent virus shedding, starting from 18 days post inoculation (d.p.i.), till 164 d.p.i. was detected, while the control animals remained negative. The virus was successfully reisolated in terrapene heart cell culture in 16 of 20 inoculated animals of both species. Histopathology of most inoculated animals revealed a lack of bone remodeling and vacuolisation in kidney tubuli which supports the described pathogenesis of nephropathy and osteodystrophy. Anti- ToPV antibody titres ranged from 1:2 to >1:256 in 13 of 20 animals, whereas all control animals were seronegative. The study proofed the Henle Koch`s postulates of ToPV as causative agent for shell dystrophy and kidney disease in both testudo species. The proposed species specific sensitivity towards clinical disease was not observed.

Mycoplasma hafezii sp. nov., isolated from the trachea of a peregrine falcon (Falco peregrinus).

Mycoplasma species are well known pathogens in avian medicine, especially in poultry. However, several Mycoplasma species have been regularly found in the respiratory tract of birds of prey which seem to be commensals in these bird species. In previous studies, an unknown Mycoplasma species which caused false positive results in a Mycoplasma meleagridis-specific PCR, was isolated from a tracheal swab of a clinically healthy, captive, adult peregrine falcon (Falco peregrinus). The isolate appeared in typical fried-egg-shaped colonies on SP4 agar plates and was dependent on sterol for growth. Acid was produced from glucose, but no arginine or urea was hydrolysed. The temperature range for growth was 28-44 °C, with an optimum at 37 °C. Strain M26T was serologically distinct from all species of the genus Mycoplasma with 16S rRNA gene sequence similarity ≥94 %. Biochemical, serological and molecular biological properties demonstrate that this organism represents a novel species of the genus Mycoplasma, for which the name Mycoplasma hafezii sp. nov. is proposed; the type strain is M26T (NCTC 13928, DSM 27652).

Lactococcus lactis, causative agent of an endocarditis valvularis and parietalis thromboticans in the allis shad, Alosa alosa (L.).

Since the 1940s, the anadromous allis shad, Alosa alosa (L.), has suffered population declines throughout its distribution range in Europe. In context of EU-LIFE projects for the reintroduction of the allis shad in the Rhine system, a comprehensive study was started in 2012 to investigate infectious diseases occurring in allis shad. In course of the study, 217 mature and young-of-the-year allis shad originating from the wild population from the Gironde-Garonne-Dordogne system (GGD-system) and the Rhine system as well as 38 allis shad from the breeding population were examined by use of bacteriological and histological methods. In 2012 and 2014, an endocarditis valvularis thromboticans caused by a coccoid bacterium was detected in 16% and 25% of mature allis shad originating from the GGD-system. Results of microbiologic examinations, including biochemical characteristics, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequence analysis, revealed Lactococcus lactis as causative agent of this infection. This is the first report of an endocarditis valvularis and parietalis thromboticans caused by Lactococcus lactis in fish. Possible sources of infection as well as the impact for the reintroduction programme are discussed.

Investigations on a cryopreservation protocol for long-term storage of psittacine spermatozoa using cockatiel semen as an example.

The aim of the present study was the establishment of an effective protocol for cryopreservation of psittacine semen. Therefore, pooled semen samples of 30 cockatiels (Nymphicus hollandicus) were diluted with modified Lake diluent (1:4), partitioned into four equal parts. Three portions were mixed with three cryoprotectants (dimethylacetamide, dimethyl sulfoxide, glycerol) in 4%, 8% and 12% final concentration, respectively, whereas the 4rth part served as control. Altogether, 96 incremental diluted semen samples were obtained for investigation. Each cryoprotective agent (CPA) in each final concentration was evaluated regarding sperm motility immediately after dilution and another four times every 30 min. Sperm viability was evaluated 0 and 120 min after dilution using the fluorescence stain SYBR® Green/propidium iodide. Sperm morphology was evaluated 0 and 120 min after dilution using eosin B stains. Glycerol demonstrated a lethal effect on cockatiel spermatozoa in all concentrations, whereas dimethylacetamide (DMA) in 8% final concentration proved to have the least adverse effect on semen parameters. Comparison of quick and slow freezing methods using DMA 8% revealed significantly higher rates of viable and motile spermatozoa after computer-controlled rate freezing. Two insemination experiments resulted in an egg fertility rate of 92.59% and 67.65% after artificial insemination with freshly collected semen samples, compared to 30.77% and 18.00% egg fertility rates using frozen/thawed semen. Altogether, 12 chicks hatched out of eggs inseminated with cryopreserved semen. To our knowledge, this is the first time for cockatiels to be successfully reproduced after artificial insemination using cryopreserved semen.

Investigation into diseases in free-ranging ring-necked pheasants (Phasianus colchicus) in northwestern Germany during population decline with special reference to infectious pathogens.

The population of ring-necked pheasants (Phasianus colchicus) is decreasing all over Germany since the years 2008/2009. Besides impacts of habitat changes caused by current rates of land conversion, climatic influences or predators, a contribution of infectious pathogens needs also to be considered. Infectious and non-infectious diseases in free-living populations of ring-necked pheasants have been scarcely investigated so far. In the present study, carcasses of 258 deceased free-ranging pheasants of different age groups, predominantly adult pheasants, collected over a period of 4 years in the states of Lower Saxony, North Rhine-Westphalia and Schleswig-Holstein, were examined pathomorphologically, parasitologically, virologically and bacteriologically, with a focus set on infectious pathogens. A periocular and perinasal dermatitis of unknown origin was present in 62.3% of the pheasants. Additional alterations included protozoal cysts in the skeletal musculature (19.0%), hepatitis (21.7%), enteritis (18.7%), gastritis (12.6%), and pneumonia (11.7%). In single cases, neoplasms (2.6%) and mycobacteriosis (1.7%) occurred. Further findings included identification of coronaviral DNA from trachea or caecal tonsils (16.8%), siadenoviral DNA (7.6%), avian metapneumoviral RNA (6.6%), and infectious bursal disease viral RNA (3.7%). Polymerase chain reaction (PCR) on herpesvirus, avian influenza virus (AIV), paramyxovirus type 1 (PMV-1), avian encephalomyelitis virus (AEV), and chlamydia were negative. Based on the present results, there is no indication of a specific pathogen as a sole cause for population decline in adult pheasants. However, an infectious disease can still not be completely excluded as it may only affect reproduction effectivity or a certain age group of pheasants (e.g., chicks) which were not presented in the study.

Seasonal and genera-specific variations in semen availability and semen characteristics in large parrots.

In large parrots electro-stimulation is suitable for collecting semen, and therefore, to facilitate semen examination and artificial insemination. Previous studies have detected differences in the semen collection success rate and semen parameters between psittacine genera. It remained unclear whether these differences were genera-related, seasonal variations or depend on the males' relationship status. To answer these questions, semen collection and spermatological analysis were performed for four psittacine groups (macaws, amazons, eclectus parrots and cockatoos) over 13 months. In one breeding facility, semen collection was attempted in 82 males using electro-stimulation twice monthly. A complete spermatological evaluation was performed on 435 semen samples. Volume, color, consistency, contamination and pH of semen, as well as motility, progressive motility, sperm concentration, total sperm count, viability, and morphology of spermatozoa were evaluated. Seasonality affected the collection success rate in macaws and amazons. Thereby, in amazons a distinct peak was observed several days before and around oviposition, whereas eclectus parrots and cockatoos produced semen all year round. The average sperm concentration was highest in eclectus parrots (2.7 × 106 sperm/µl) and lowest in macaws (35.6 × 103 sperm/µl). The differences in the semen collection success rate and semen parameters seem to coincide with the bird's breeding biology. The collected data allows a prognostic estimation when semen collection seems favorable, and may be taken as orientation values for semen analysis in these species.

First report of a cystic malformation on the upper jaw of hatchery-reared allis shad Alosa alosa.

The anadromous allis shad Alosa alosa has suffered dramatic population declines throughout Europe and is currently considered as endangered throughout its entire distribution range. In order to reestablish allis shad in the River Rhine, which formerly housed one of the largest and most important populations, an EU-LIFE Project 'The re-introduction of allis shad in the Rhine system' was started in 2007. In course of the LIFE+ Projects, allis shad larvae bred from genitor fish of the Gironde-Garonne-Dordogne population in France were reared in a pilot ex situ stock plant pilot facility in Aßlar, Germany. At an age of 1-2 months, about 100% of these fish developed approximately 0.5- to 0.8-cm large, fluid-filled, transparent cysts in conjunction with the upper jaw. The performed microbiological, virological, parasitological and histological examinations did not detect any infectious agents. Possible causative agents are discussed with regard to environmental factors and the nutrition of larvae. In conclusion, the observed malformations are considered a sign for a severe health problem and therefore a risk for the successful breeding of allis shad in aquaculture.

The effect of endoscopic sterilization on reproductive behavior and pair bond maintenance of feral pigeons (Columba livia).

OBJECTIVE: Problems related to feral pigeons (Columba livia) in cities mainly result from their large numbers due to uncontrolled population growth. The aim of this study was to evaluate whether endoscopic guided sterilization affects the reproductive behavior of feral pigeons under experimental conditions, with the intention of assessing this technique as a potential method for feral pigeon population control. MATERIALS AND METHODS: Five groups of four pairs of feral pigeons each were studied from 8 weeks before, to 7 weeks after sterilization. Both the male and female of the first pair of each group were sterilized, in the second pair only the female and in the third pair only the male was sterilized. The fourth pair acted as a control. All eggs laid were candled to assess fertility. RESULTS: Surgical sterilization had minimal effects on behavior and therefore seems not to have impact on possible field application for population control. All pairs maintained their pair bonds and continued to defend their nesting sites against other pigeons. Only one female copulated with a foreign fertile male while her primary partner was debilitated due to surgery, but returned to him as soon as he recovered. All eggs laid more than 5 days after male sterilization were infertile, whereas all control pairs had fertile eggs. Only one fertile clutch was produced, 5 days after the male's sterilization. Therefore it is assumed that males remain fertile for a limited period of time. CONCLUSION: Endoscopic sterilization seems to be a promising method for field control of feral pigeon populations and sterilization of the male only seems sufficient.

[Role of the poultry red mite (Dermanyssus gallinae) in the transmission of avian influenza A virus]. / Rolle der Roten Vogelmilbe (Dermanyssus gallinae) bei der Übertragung von aviärem Influenza-A-Virus.

OBJECTIVE: The aim of this study was to investigate the role of the poultry red mite (Dermanyssus [D.] gallinae) in the horizontal transmission of avian influenza A virus (AIV) to chickens. This mite is the most common ectoparasite in poultry worldwide, and may play a role in the spread of infectious agents including AIV. Currently, the control of mites is difficult due to frequently developed resistance to many acaricides, their nocturnality and their ability to survive hidden without feeding for months. MATERIALS AND METHODS: D. gallinae were collected in a commercial layer farm and housed in self-made fibreboard boxes. SPF chickens were intravenously infected with AIV strain A/turkey/Ontario/7732/1966 (H5N9). The viraemia in chickens was monitored and at an appropriate time point about 1000 mites were allowed to suck on the AIV infected chickens. Re-isolation of the virus from blood-filled mites was tried daily for 14 days using chicken embryo fibroblast cultures and embryonated chicken eggs. Subsequently, the virus containing mites were placed into boxes that contained naïve SPF chickens to enable virus transmission from mites to chickens. Possible transmission to the chickens was examined using clinical signs, serology, gross lesions, histopathology and immunohistochemistry. RESULTS: Chickens developed a dose-dependent viraemia one day after infection, therefore this day was chosen for the bloodmeal of the mites. AIV was detected in mites after bloodsucking on AIV-infected chickens over a 10-day period. Naïve SPF chickens were infected during bloodsucking of AIV carrying mites. AIV isolates in mites and in chickens were undistinguishable from the original AIV inoculum by RT-PCR. CONCLUSIONS: D. gallinae ingested AIV during bloodmeals on AIV infected chickens and are able to transmit AIV to SPF chickens. Therefore, mites serve as mechanical vector of AIV and may play a major role in the circulation of AIV within a facility or area although the life span of infectious virus in the mite is limited. CLINICAL RELEVANCE: The proven transmission requires more than ever a systematic control of this ectoparasite in order to maintain poultry health and productivity. The demonstrated vector function of this mite is of great significance for poultry flocks all over the world.

Toltrazuril does not show an effect against pigeon protozoal encephalitis.

The protozoan parasite Sarcocystis calchasi causes a severe neurologic disease in domestic pigeons (Columba livia f. dom.) named pigeon protozoal encephalitis. Recently, the parasite has also been reported in psittacines causing a virtually identical disease with fatal outcome. So far, an etiological treatment of S. calchasi infections in pigeons or psittacines is unknown. The present study evaluates the effectiveness of the anticoccidian drug toltrazuril against S. calchasi and the influence of the timepoint of treatment. Therefore, nine domestic pigeons were inoculated with 400 S. calchasi sporocysts and treated with toltrazuril (25 mg/kg) in groups of three pigeons each at dpi 10/11 and dpi 40/41 and on two consecutive days at the onset of neurologic signs. After euthanasia at dpi 73, tissue samples including brain and skeletal muscles were examined by histology and S. calchasi-specific real-time PCR. All pigeons independent of the group developed neurologic signs from dpi 49 onwards. Histology identified sarcocysts in the skeletal muscles and a granulomatous encephalitis in the brains. The relative amount of S. calchasi DNA was on a comparable level in all pigeons. Consequently, toltrazuril was demonstrated to be not effective against S. calchasi with the applied treatment regime. Longer treatment periods or agents other the toltrazuril may be considered for further investigations. So far, preventive measures like roofing of aviaries for prevention of infection and regular disinfection remain the most important factor in the control of S. calchasi infections.

Endoscopic assisted orchiectomy in Herman's tortoises (Testudo hermanni sp.).

OBJECTIVE: Male tortoises in captivity are often aggressive against other males or females, in particular during mating season related to hormonal influences (testosterone). Castration in males is the treatment of choice in many vertebrate species. A novel technique of minimal invasive castration is presented for Herman's tortoises (Testudo hermanni). MATERIAL AND METHODS: The procedure was performed in ten mature males. An endoscope (2.7 mm/30° angled) was inserted into a prefemoral incision on each side. The testicle was resected after ligation of the gubernaculum testis and the spermatic cord using hemoclips. RESULTS: Each testicle could be removed in approximately 20 minutes. Complications like hemorrhage or damage of adjacent tissue did not occur and all animals recovered uneventfully. CONCLUSIONS AND CLINICAL RELEVANCE: Using the bilateral prefemoral entrance offers an alternative option for orchiectomy in Herman's tortoises without dissection of the shell. Thus complications like hemorrhage or impaired wound healing followed by sequestration of the bone flap are prevented. Because of the anatomical settings and the risk of tissue damage and time consumption, a unilateral approach is not recommended. Fasting the animals is necessary due to the voluminous gastrointestinal tract of this herbivorous tortoise and emptying the urinary bladder provides more space for manipulations in the coelomic cavity and prevents clipping and cutting of adjacent organs. The magnification via the endoscope is beneficial for orientation in the coelomic cavity, which is formed by the extremely convex carapace. Without this equipment it is challenging to visualize the dorsocaudally located gonads through the relatively small incision and the inserted instruments might block the view at the surgical field. Administration of hemoclips achieved a good hemostasis and the testicles could be resected without major blood loss. The described technique is a gentle method for resection of the testicles in this species and can be adapted to other European tortoise species of equal size.

[Investigation of the practical use of a vaccination device ("Pullet Vaccinator") for young layers]. / Untersuchungen zum Einsatz des Impfautomaten "Pullet Vaccinator" in der Junghennenaufzucht.

OBJECTIVE: To evaluate the safety of layer vaccination of the vaccination device "Pullet Vaccinator", its publicised increased operational safety and the practicality of the device using serological monitoring of the vaccination success. MATERIALS AND METHODS: In a first trial, two veterinarians experienced in the syringe method vaccinated hens using the syringe and the vaccination device, respectively. After 1 hour, the equipment was switched and both veterinarians continued to vaccinate for a further hour. The second trial proceeded as in the first trial, except with untrained persons. For each of the four vaccination groups (experienced/syringe; experienced/device; untrained/syringe; untrained/device), the number of vaccinated hens was counted and 20 hens were dissected from each group to compare the impact of both inoculation methods on the animals. To monitor vaccination success, blood was collected for serological screening. On the final blood collection day, further hens per group were dissected to evaluate possible long-term injuries. RESULTS: The vaccination device offers greater occupational safety compared to the syringe method. Vaccination injuries to the hens' breast muscles were more pronounced with the syringe application. For experienced persons, the number of vaccinated animals per hour was approximately doubled using the syringe compared to the device. For the untrained, a comparable number of vaccinated animals was recorded for both methods. Serological monitoring did not show any significant differences in antibody response to the vaccination between both methods. CONCLUSION AND CLINICAL RELEVANCE: In certain points, the device proved technically imperfect and should be revised for improved use in the field.

Soft plastron, soft carapace with skeletal abnormality in juvenile tortoises. Histopathology and isolation of a novel picornavirus from Testudo graeca and Geochelone elegans.

OBJECTIVE: A disease is described in juvenile tortoises (Testudo graeca and Geochelone elegans) consisting mainly of a soft carapace, soft plastron and deformed skeleton. The aim of this study was to determine histopathological lesions and the biological properties of the isolated viruses. MATERIALS AND METHODS: Clinical signs and gross pathology were determined on diseased and healthy appearing tortoises. Paraffin sections were stained with HE, PAS and Prussian Blue and histologically examined. Terrapene heart (TH-1) cell cultures served for virus isolations from 64 tissues and 104 swabs. One isolate (isolate 1243/37 tongue) was used in neutralization tests on 19 sera. RESULTS: Retarded growth and increasingly soft plastron and carapace were the prominent signs in diseased tortoises. Pathological lesions consisted of dilated urinary sac, enlarged kidneys and livers. Histopathologically, hepatic hemosiderosis, hypoplastic anaemia, congestive glomerulonephrosis and osteodystrophy were seen. A novel vi- rus ("virus X") was isolated from 64 organs and 79 of 104 swabs. The isolated viruses were identified as a novel chelonid picornavirus based on cytopathic effect, resistance to chloroform and stability at low pH. Co-cultivation with 5-iodo-2'-deoxyuridine and actinomycin D did not reduce virus titres. Electron microscopically, round, non-enveloped particles (25-30 nm) were detected. Neutralizing antibodies to the isolate 1243/37tongue were present in 17 of 19 sera from seven species of tortoises. CONCLUSION AND CLINICAL RELEVANCE: Nephropathy, osteodystrophy and virus isolations suggest a viral aetiology. Metabolic bone disease is the major differential diagnosis. Further investigations in vivo are needed to evaluate the likely effects of the picornavirus on tortoises.

Comparison of conventional and computer-assisted semen analysis in cockatiels (Nymphicus hollandicus) and evaluation of different insemination dosages for artificial insemination.

Many psittacine species are threatened in the wild and also rare in captivity. Therefore, successful conservation and breeding programs are important to save these species. Unfortunately, clutches in conservation programs are frequently infertile. Semen evaluation is beneficial to investigate the causes of infertility and is advisable before artificial insemination (AI). In this study, we analyzed the semen of cockatiels (Nymphicus hollandicus) using two different methods and investigated different insemination dosages for AI. Cockatiels (n = 30) were divided into two groups (group A: nine males; group B: six males). The males in group B were endoscopically sterilized, whereas the males in group A were used as semen donors. In the first part of the study, the semen of males in group A was evaluated by semen analysis. Semen samples were collected by the massage technique and examined using a conventional light microscope and a computer-assisted semen analyzer for comparison. Results demonstrated that the evaluations of motility, progressive motility, and sperm concentration, but not of live/dead ratio, correlated strongly for both methods. However, the results for sperm concentration, progressive motility, and live/dead ratio differed significantly. In the second part of our study, the volume and quantity of spermatozoa of the semen samples were adjusted and used for AI of females of group B. Intravaginal insemination with 250,000 spermatozoa resulted in five of 17 (29%) eggs fertilized; however, intracloacal insemination resulted in only four of 57 (7%) eggs fertilized at 232,000 and 250,000 spermatozoa but none at higher or lower dosages.

[Monitoring the success of veterinary treatment in rehabilitated and released birds of prey using radiotelemetry]. / Monitoring rehabilitierter und ausgewilderter Greifvögel mittels Telemetrie als Erfolgskontrolle.

OBJECTIVE: Free-ranging birds of prey brought to veterinary practice should only be treated after thorough diagnostics. Before their release back into the wild, specific training - including falconry techniques - may be necessary, depending on raptor species and age. Rehabilitated birds of prey were monitored using radiotelemetry after release back into the wild. The success of veterinary therapy and the prognosis of treated diseases/injuries in free-ranging birds were evaluated. In addition, the use of radiotelemetry as a simple technique for surveillance was evaluated. The project was undertaken in cooperation with schools as a contribution to environmental education. MATERIAL UND METHODS: Three common buzzards (Buteo buteo) and one kestrel (Falco tinnunculus)were treated and released with a radio transmitter attached to a tail feather. They were tracked daily (by car or plane), observed using binoculars and their GPS coordinates were documented. RESULTS: One transmitter was lost early, making monitoring of the bird impossible. Three birds were monitored over a period of more than 14 days. These birds were successfully reintroduced into the wild, as documented from courtship displays and mating. The longest flight distance achieved was 44 km. CONCLUSIONS: Veterinary treatment aimed at rehabilitating feral birds can be successful. Radiotelemetry is a suitable tool to monitor free-ranging birds. The application of this technique is performed readily by laypeople (school students). CLINICAL RELEVANCE: Being in agreement with other studies, this data should motivate veterinarians to treat wild birds of prey for rehabilitation.

Comparison of diagnostic tools for the detection of aspergillosis in blood samples of experimentally infected falcons.

Antemortem diagnosis of avian aspergillosis is very challenging. Diagnostic assays using blood samples would aid in an early and more definitive diagnosis. In the current study, detection of anti-Aspergillus antibodies, Aspergillus antigen, and Aspergillus toxin (fumigaclavine A), protein electrophoresis and measurement of acute-phase protein concentrations were performed on serum of 18 adult and plasma of 21 juvenile gyr-saker hybrid falcons (Falco rusticolus x Falco cherrug). Adult (n = 15) and juvenile (n = 18) falcons were experimentally inoculated with different dosages of the same strain of Aspergillus fumigatus and an additional three falcons from each age group were used as uninfected control animals. Blood samples were collected prior to inoculation and at 28 days postinoculation. Of the 33 inoculated falcons, 16 demonstrated clinical signs (vomiting, greenish urates, dyspnea, ruffled feathers) commonly associated with aspergillosis and in 14 falcons necropsy revealed aspergillosis granulomas confirmed by mycology and histopathology. Positive galactomannan results were rare, with only 3/15 positive samples from adult falcons and none in the juvenile birds. Most of the inoculated falcons showed an increase of serum amyloid A (66.7%) and haptoglobin (70.4%), but fumigaclavine A was not detected in the blood from any of the experimental animals. Elevated antibody indices were detected in 96.7% of the inoculated birds, but also in 66.7% of the controls. Significant decreases in albumin:globulin ratio were obvious in 81.5% of the inoculated birds, including 100% of the birds with granulomas. Blood from falcons with granulomas demonstrated significantly increased concentration values of alpha 2 and ß globulins, decreased percentages of prealbumin and albumin, and increased percentages of alpha 2 and ß globulins compared to inoculated falcons without granulomas. In conclusion, acute-phase proteins and the electrophoretic profile of birds challenged with A. fulmigatus show significant alterations, which in combination with other diagnostic procedures, assist in the early diagnosis of avian aspergillosis.