Uptake of Neonicotinoid Insecticides by Water-Foraging Honey Bees (Hymenoptera: Apidae) Through Guttation Fluid of Winter Oilseed Rape.

The water-foraging activity of honey bees (Apis mellifera L.) on guttation fluid of seed-coated crops, such as winter oilseed rape (WOR; Brassica napus L.), has not yet been evaluated. We analyzed the uptake of active substances (a.s.) in guttation fluid by evaluating residues of honey-sac contents. In autumn, insecticide residues of up to 130 µg a.s. per liter were released in WOR guttation fluid; this concentration is noticeably lower than levels reported in guttation fluid of seed-coated maize. Until winter dormancy, the concentrations declined to <30 µg a.s. per liter. In spring, residues were linked to prewintered plants and declined steadily until flowering. The maximum release of residues in guttation fluid of seed-coated WOR occurs on the first leaves in autumn when the colonies' water demand decreases. For the first time, proof for the uptake of guttation fluid from seed-coated WOR by honey bees was provided by measuring residues in individual honey-sac contents. In total, 38 out of 204 samples (19%) showed residues of thiamethoxam at concentrations ranging from 0.3 to 0.95 µg per liter while the corresponding concentrations in guttation fluid of WOR varied between 3.6 to 12.9 µg thiamethoxam per liter. The amounts of thiamethoxam we found in the honey sacs of water-foraging honey bees were therefore below the thresholds in nectar and pollen that are considered to have negative effects on honey bees after chronic exposure.

Benign and Malignant Proliferative Fibro-osseous and Osseous Lesions of the Oral Cavity of Dogs.

Ossifying fibroma (OF) and fibrous dysplasia (FD) are benign, intraosseous, proliferative fibro-osseous lesions (PFOLs) characterized by replacement of normal bone by a fibrous matrix with various degrees of mineralization and ossification. Osteomas are benign tumors composed of mature, well-differentiated bone. Clinical, imaging, and histologic features of 15 initially diagnosed benign PFOLs and osteomas of the canine oral cavity were evaluated. Final diagnoses after reevaluation were as follows: OF (3 cases), FD (4 cases), low-grade osteosarcoma (LG-OSA) (3 cases), and osteoma (5 cases). Histology alone often did not result in a definitive diagnosis for PFOL. OF appeared as a well-circumscribed, radiopaque mass with some degree of bone lysis on imaging. Most lesions of FD showed soft tissue opacity with bone lysis and ill-defined margins. Low-grade OSA appeared as a lytic lesion with a mixed opacity and ill-defined margins. Osteomas were characterized by a mineralized, expansile, well-circumscribed lesion. Although histologic features of PFOLs were typically bland, the lesions diagnosed as LG-OSA had some features of malignancy (eg, bone invasion or a higher mitotic index). Treatment varied widely. Of the 10 dogs with benign PFOL or osteoma with known outcome (10/12), 9 showed either complete response (6/10) or stable disease (3/10) after treatment. Of the 2 dogs with LG-OSA with known outcome, 1 showed complete response after curative intent surgery, but 1 patient had recurrence after partial maxillectomy. Definitive diagnosis of mandibular/maxillary PFOL is challenging via histopathologic examination alone, and accurate diagnosis is best achieved through assimilation of clinical, imaging, and histopathologic features.

Early tumor response to intraarterial or intravenous administration of carboplatin to treat naturally occurring lower urinary tract carcinoma in dogs.

BACKGROUND: Survival times and tumor responses associated with malignant neoplasia of the lower urinary tract are poor despite the vast array of current treatments. Therefore, the evaluation of alternative treatments, such as intraarterial administration of chemotherapy (IAC) should be considered. OBJECTIVE: To describe a technique for superselective catheterization for IAC and to evaluate initial tumor response by ultrasonography after both IAC and intravenous administration of chemotherapy (IVC). ANIMALS: Client-owned dogs with lower urinary tract neoplasia treated with either IVC (n = 15) or IAC (n = 11). METHODS: Retrospective study. An arterial approach via the carotid or femoral artery was utilized to obtain superselective access and administer chemotherapy in the IAC cases. Medical record review was performed, data were recorded, and recorded variables were evaluated statistically. RESULTS: Intraarterial chemotherapy was successfully administered in all cases. There was a significantly greater decrease in longest unidimensional measurement in the IAC group as compared to the IVC group (P = .013). The IAC group was also significantly more likely to have a tumor response as assessed by modified RECIST guidelines (P = .049). Dogs in the IAC group were significantly less likely to develop anemia (P = .001), lethargy (P = .010) and anorexia (P = .024). CONCLUSION AND CLINICAL IMPORTANCE: This study demonstrated the feasibility and efficacy of performing IAC for lower urinary tract neoplasia. Further investigation is necessary as the follow-up time was short and the impact on long-term outcome and survival was not determined.

Signalment, clinical presentation, concurrent diseases, and diagnostic findings in 28 dogs with dynamic pharyngeal collapse (2008-2013).

BACKGROUND: Most information about pharyngeal collapse in dogs is anecdotal and extrapolated from human medicine. A single case report describing dynamic pharyngeal collapse in a cat has been published, but there is no literature describing this disease process in dogs. OBJECTIVE: To describe the signalment, clinical presentation, concurrent disease processes, and imaging findings of a population of client-owned dogs with pharyngeal collapse. ANIMALS: Twenty-eight client-owned dogs with pharyngeal collapse. METHODS: Radiology reports of dogs for which fluoroscopy of the respiratory system was performed were reviewed retrospectively. Patients with a fluoroscopic diagnosis of pharyngeal collapse were included in the study population. Data regarding clinical signs, diagnostic, and pathologic findings were evaluated. RESULTS: Twenty-eight dogs met the inclusion criteria. The median age of affected patients was 6.6 years, whereas median body condition score was 7/9. The most common clinical signs were coughing (n = 20) and stertor (n = 5). In 27 of 28 cases, a concurrent or previously diagnosed cardiopulmonary disorder was detected. The most common concurrent disease processes were mainstem bronchi collapse (n = 18), tracheal collapse (n = 17), and brachycephalic airway syndrome (n = 8). Fluoroscopy identified complete pharyngeal collapse in 20 of 28 dogs. CONCLUSIONS: Pharyngeal collapse is a complex disease process that likely is secondary to long-term negative pressure gradients and anatomic and functional abnormalities. Based on the findings of this study, pharyngeal fluoroscopy may be useful diagnostic test in patients with suspected tracheal and mainstem bronchial collapse to identify concurrent pharyngeal collapse.

An ORF1-rearranged hepatitis E virus derived from a chronically infected patient efficiently replicates in cell culture.

Hepatitis E is an increasingly reported disease in industrialized countries. Studies on the replication cycle of hepatitis E virus (HEV) are hampered due to the lack of efficient and robust cell culture systems for this virus. We describe the successful isolation of HEV derived from a chronically infected kidney transplant patient held under immunosuppressive therapy. Inoculation of serum sample 47832 onto the human lung carcinoma cell line A549 resulted in the replication of the virus as shown by RT-qPCR. This novel human-derived HEV strain is closely related to a wild boar-derived genotype 3 strain, which did not replicate in A549 cells. It carries a 186 nucleotide insertion in the hypervariable ORF1-region, derived from two parts of its ORF1. By passaging of the infected cells, a cell line continuously producing HEV particles was generated as demonstrated by RT-qPCR, immuno-electron microscopy, density gradient centrifugation and immunohistochemistry. Replication of the produced virus was demonstrated after its inoculation onto fresh A549 cells and two consecutive passages, whereas heating at 65 °C for 2 min abolished its infectivity. Several point mutations scattered along the whole genome were present in the HEV strain from the second passage; however, the ORF1 insertion was still present. Previously, cell culture isolation of two other HEV strains carrying insertions in their hypervariable regions, but originating from human ribosomal protein genes, has been described. The findings may indicate that cell culture adaptation of is mostly dependent on the length and position of the insertion, rather than from the sequence itself.

Replication of hepatitis E virus in three-dimensional cell culture.

Hepatitis E is an acute, viral hepatitis epidemic in developing regions, but which is detected with increasing frequency in sporadic form in developed regions. Pigs and possibly some other mammals are considered reservoirs of zoonotic infection with hepatitis E virus (HEV). However, whilst the relative significance of potential transmission routes from pigs to people is still unclear, the consumption of raw or undercooked pig meat has been implicated as a source of HEV infection. The lack of information about HEV zoonotic transmission is due in part to the difficulties of in vitro propagation of HEV. The Rotating Wall Vessel (RVW) has been described as a useful tool for the culture of cell lines in a 3-dimensional (3D) configuration. The aim of this work was to develop a 3D cell culture system for HEV to facilitate studies into the viability of virions contaminating pig tissues. This study, demonstrated that HEV can replicate efficiently in the RWV in human hepatoblastoma PLC/PRF/5 cells for up to 5 months not only by real time RT-PCR but also by detection of complete virions via electron microscopy. Furthermore, the replication of HEV progeny was observed by detecting HEV RNA by RT-PCR. The progeny were able to infect fresh 3D cultures, showing that this method is able to produce infectious hepatitis E virions.

Reliability of T2-weighted sagittal magnetic resonance images for determining the location of compressive disk herniation in dogs.

Magnetic resonance imaging is used commonly to diagnose intervertebral disk herniation in dogs. It is common to locate areas of suspected compression on sagittal T2-weighted (T2-W) images and then obtain limited transverse images in these areas to reduce the acquisition time (a step-by-step approach). Our objective was to assess the frequency of correct localization of spinal cord compression due to disk herniation using only the sagittal images. The results from isolated readings of the sagittal T2-W images alone or combined with a single-shot fast spin echo (SSFSE) slab in 118 dogs were compared with a gold standard, based on a consensual reading of all images available, including complete transverse images across the entire spinal segments under study. The sites of compression were localized correctly from the sagittal images in 89.8% of dogs. If only the most significant lesions were accounted for, the percentage increased up to 95.2%. In 54.9% of the readings with incorrect localization, the actual compressive site was immediately adjacent to the one suspected from review of the sagittal images. The frequency of correct localization was higher in the cervical region, and was increased by examination of the SSFSE slab. The most common cause of disagreement was the presence of multiple degenerate bulging disks. Based on these results we recommend obtaining transverse images across the entire segment when multiple bulging disks are present. It is also recommended to obtain transverse images across the spaces immediately adjacent to the suspected site of herniation from review of the sagittal images.

Complication rate, outcome, and risk factors associated with surgical repair of fractures of the lateral aspect of the humeral condyle in dogs.

OBJECTIVE: To determine outcome, report complications, and risk factors for complications associated with the surgical treatment of fractures of the lateral aspect of the humeral condyle (FLHC) in dogs. STUDY DESIGN: Fifty dogs with FLHC that had been repaired surgically were included in this retrospective case series. METHODS: The medical records and radiographs from 50 dogs with FLHC from 1990--2006 were analyzed. RESULTS: The complication rate was 30%. For every 30-minute increase in surgical time the odds of a complication increased by 2.00 (95% CI: 1.06, 3.77). For every 10 degrees increase in the angle between the transcondylar screw and a line between the epicondyles viewed on a craniocaudal radiograph there was a 4.82 (95% CI: 1.23, 18.86) greater odds of an intracondylar fracture gap appearing on postoperative radiographs. When an intracondylar fracture gap was present after surgery there was a 25% long-term radiographic outcome failure rate, whereas there was a 0% radiographic failure rate in the dogs in which an intracondylar fracture gap was not present. In conclusion, transcondylar screw placement parallel to the epicondylar line should be attempted for the repair of FLHC. An increase in surgical time leads to an increase in postoperative complication rate. The presence of an intracondylar fracture gap may be associated with fracture failure. CLINICAL SIGNIFICANCE: Screw angulation predisposed to postoperative intracondylar fracture gap which in turn may be associated with higher fracture fixation failure rate. The dogs that had prolonged surgical times had an increased complication rate.

Isolation, identification and characterization of group A rotavirus from a chicken: the inner capsid protein sequence shows only a distant phylogenetic relationship to most other avian group A rotaviruses.

Rotavirus particles were identified in the intestinal content of a 35-day-old stunted chicken. The virus was isolated, RNA pattern was analysed and the viral genome segment 6 was sequenced. In particular, the sequence data showed a very close similarity to the chicken rotavirus isolate Ch-1 (99.2% amino acid homology), this is distantly related to all known avian rotaviruses and supports the existence of different VP6 types amongst avian group A rotaviruses.

[Severe course of ornithosis and its intensive care and diagnostic problems--a case report]. / Schwerer Verlauf einer Ornithose und ihre intensivmedizinische und diagnostische Problematik--eine Kasuistik.

Ornithosis is still a life-threatening illness unless diagnosed early and adequately treated with antibiotics. Diagnosis, however, is difficult because often no direct contact with birds is obvious. We report on the course of a severe case of ornithosis in a 45-year-old man who had no previous health problems. Infection with Chlamydia psittaci probably occurred by inhaling of contaminated dust produced by flocks of wild pigeons. A special feature of the case is that commonly used serological tests (complement fixation reaction, CFR; ELISA) failed to yield clear results. Since propagation of Chlamydia psittaci requires special biological laboratory containment facility no. 3, we were able to detect the pathogen in faeces, blood, and urine using the direct immunofluorescence antibody technique (DIFT) and a modified peroxidase antiperoxidase technique (PAP). A lot of Chlamydia psittaci was found. These results were verified by electron microscopy. As a result, the treatment could be quickly adjusted to the application of doxycyclin, which led to a rapid improvement in the patient's condition. This direct detection of Chlamydia psittaci by DIFT, the PAP technique or electron microscopy, also in faeces, blood and urine specimens, represents a useful addition to the usual microbiological methods for the diagnosis of this pathogen.

[Endomycoses--a problem in the diagnosis of Chlamydia psittaci infections in chickens]. / Endomykosen--ein Problem bei der Diagnostik von Chlamydia-psittaci-Infektionen bei Hühnern.

Two hens, which died at an age of 42 respectively 45 weeks, showed firstly lesions of endomycosis as sole cause of illness or death at necropsy and in light microscopic investigations. With peroxidase-antiperoxidase-(PAP-)technique and in electron microscopic investigations, however, Chlamydia psittaci were also found in very large numbers in both chickens, and occurred here particularly in lungs, liver, kidneys, spleen, myocard and ovary. The results showed, that endomycosis and chlamydial infections can occur in chickens simultaneously. In such cases of mixed infections the PAP-technique may be very helpful for rapid and reliable detection of Chlamydia psittaci in formalin-fixed and paraffin-embedded chicken tissues.

[The detection of Toxoplasma gondii in abortion tissues of sheep using the polymerase chain reaction]. / Der Nachweis von Toxoplasma gondii in Abortgeweben vom Schaf mittels der Polymerase-Kettenreaktion.

A polymerase chain reaction was applied to detect Toxoplasma gondii DNA in placental and fetal tissue samples of 47 unselected ovine abortions of the lambing season 1990/91 (Baden-Württemberg, Rhineland, Hesse). For the amplification a 190 bp or 223 bp sequence of the B1-gene of T. gondii was selected as the target sequence. Both sequences were detected in five abortions. All positive results were immunohistochemically confirmed using the peroxidase antiperoxidase technique (PAP-staining). Thus, in Germany, too, T. gondii infection in sheep during pregnancy should be considered as a possible cause of abortions, particularly in case of abortions of unknown genesis.

[Natural transmission of microsporidia (Encephalitozoon cuniculi) by way of the chicken egg]. / Natürliche Ubertragung von Mikrosporidien (Encephalitozoon cuniculi) über das Hühnerei.

100 chicken embryos were examined for naturally occurring infections with Encephalitozoon cuniculi (E.c.). Two embryos were found to be dead, the others were killed at an age of 18 days. Samples from the oesophagus, intestine, liver, kidneys, heart and brain were collected and examined by immunohistochemical methods. E.c. was found in about 40% of the embryos. This microsporidian was observed in all organs, particularly, however, in the oesophagus, intestine, brain and heart. The number of E.c. was significantly larger in the two embryos that died than in the others. The investigations demonstrated, that E.c. may be transmitted naturally to the chicken egg. Further, the results suggested that E.c. may be a cause of death in chicken embryos. In most cases, however, E.c. infections are inapparent in embryos. Such inapparent infections may be an important way of contamination of chicken production units with E.c.

[Naturally-acquired microsporidia (Encephalitozoon cuniculi) infections in hens]. / Natürliche Mikrosporidien-(Encephalitozoon-cuniculi-)Infektionen bei Hühnern.

During a short period of time, four approximately ten months old chickens from an egg production (group A) unit developed symptoms of apathy, lameness, mild diarrhoea and loss of weight. The layers were killed and submitted for necropsy. For the first time, microsporidia infection, probably Encephalitozoon cuniculi (E.c.) was diagnosed in chickens. The diagnosis was mainly based on immunohistochemical methods. E.c. was found particularly in the cells of the cutaneous mucous membrane of the crop and in epithelial cells, the cells of the propria mucosae and the smooth muscle cells of the small intestine. E.c. also frequently occurred in the fibres of the skeletal muscle. Intracellularly, E.c. and its development stages were located primarily in the cytoplasm and less frequently in the nucleus of cells. Retrospective investigations on tissue samples from a domestic cock and 9 hens by immunohistochemical methods also revealed occurrence of this microsporidian. The results showed that in chickens E.c. infections can cause multi-symptomatic illness but may also be clinically inapparent.