Case of suspected theobromine poisoning in dairy cattle

High-yielding dairy cows collapsed and died displaying signs of a disturbed central nervous system (muscle tremor, convulsion) and a considerably reduced body condition score. An intense diagnostic screening did not allow to confirm a definite diagnosis. Therefore, further analyses including an evaluation of feeds and feeding were initiated. The herd was fed a total mixed ration (TMR) based on corn and grass silage of moderate nutritive value supplemented with various amounts of chocolate chips. In retrospect, the amount of chocolate chips added to the respective TMR batches could not be quantified. These chips were purchased from a company producing bakery and chocolate products for human consumption and added to the silage and therefore to the TMR in order to increase the energy intake of the animals. Because the TMR, which was fed during the time of the incidence, was no longer available, a sample of a later batch of silage was examined. The amount of chocolate chips was quantified (0.44% per fresh matter), and a theobromine analysis was performed in the chocolate chips of the available batch (69.7 mg/100 g fresh matter). Because of the possible link between chocolate intake and observed signs, an immediate cessation of using the chocolate chips was recommended in addition to an optimisation of the TMR, that is an increase of the energy density. Even though the theobromine intake during the time of the incidence is unknown and information about toxicity of theobromine in ruminants is limited, we suspect that the feeding of chocolate in this case caused all signs including the sudden death of the cows. Further reasons are that no differential diagnoses were established and the problems at the farm stopped after removing the chocolate from the TMR.     

Expression of porcine endogenous retroviruses (PERVs) in melanomas of Munich miniature swine (MMS) Troll

Porcine endogenous retroviruses (PERVs) are integrated in the genome of all pig breeds. Since some of them are able to infect human cells, they might represent a risk for xenotransplantation using pig cells or organs. However, the expression and biological role of PERVs in healthy pigs as well as in porcine tumours is largely unknown. Since we and others have recently shown overexpression of a human endogenous retrovirus, HERV-K, in human melanomas, we studied the expression of PERVs in melanomas of selectively bred Munich miniature swine (MMS) Troll. This breeding herd of MMS Troll is characterised by a high prevalence of melanomas, which histologically resemble various types of cutaneous melanomas in humans. Several genetic factors have been defined when studying inheritance of melanomas and melanocytic nevi in MMS Troll. Here we show that the polytropic PERV-A and PERV-B as well as the ecotropic PERV-C are present in the genome of all melanoma bearing MMS Troll investigated. Most interestingly, in the spleen, but not in other organs, recombinant PERV-A/C proviruses were found. PERV expression was found elevated in melanomas when compared to normal skin and viral proteins were expressed in melanomas and pulmonary metastasis-derived melanoma cell cultures. During passaging of these cells in vitro the expression of PERV mRNA and protein increased and virus particles were released as shown by RT activity in the supernatant and by electron microscopy. Genomic RNA of PERV-A, -B and -C were found in pelleted virus particles. Although PERV expression was elevated in melanomas and pulmonary metastasis-derived cell cultures, the function of the virus in tumour development is still unclear.     

Molecular analysis of the Arbuscular mycorrhiza symbiosis

The molecular analysis of the arbuscular mycorrhizal (AM) symbiosis started at the beginning of this decade. The paper summarises the work of the ‘Laboratorium für Molekulare Genetik arbuskulärer Mykorrhiza’ at the MPI für terrestrische Mikrobiologie in Marburg on three major topics. Firstly, the plant response to the mycorrhizal colonisation was analysed using both, targeted and non‐targeted approaches. As an example, the localisation of the gst1 mRNA in potato mycorrhiza is shown. Second, molecular techniques were established to analyse gene expression of the fungal partner of the symbiosis. We present a differential RNA display analysis of spore germination in two AM fungi and the cloning of a gene from Glomus mosseae that shows expression at all stages of the fungal life cycle. In the last part, we introduce the work we are carrying out with a new root endophytic fungus, Piriformospora indica. Infection experiments on maize showed that despite the fungus performs as a root necrotroph, it has a positive effect on plant growth.     

Development and evaluation of an enzyme-linked immunosorbent assay for the serodiagnosis of pythiosis in dogs

Pythiosis (caused by the aquatic oomycete Pythium insidiosum) is a devastating and often fatal cause of either severe transmural gastroenteritis or locally invasive subcutaneous disease in dogs living in the southeastern United States. Although early diagnosis is essential for successful treatment, tools available for this task are limited. Therefore, we developed and evaluated an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-P insidiosum antibodies in canine serum. A soluble mycelial extract of P insidiosum was utilized as antigen in the ELISA, which was used to evaluate serum from 43 dogs with pythiosis, 8 dogs with lagenidiosis (another canine oomycosis), 16 dogs with nonoomycotic fungal or algal infections, 22 dogs with nonfungal gastrointestinal or skin disease, and 55 healthy dogs. Results were expressed as percent positivity (PP) relative to a strong positive control serum run on each plate. Medians and ranges for each of the 5 groups were as follows: pythiosis (81.7%, 50.6-98.5%), lagenidiosis (17.3%, 11.3-29.2%), other fungal or algal infections (8.2%, 4.7-15.4%), nonfungal gastrointestinal or skin disease (6.2%, 3.9-20.7%), and healthy dogs (6.7%, 3.0-15.2%). When using a cutoff value of 40% PP, the sensitivity and specificity of the ELISA both were 100%. In addition, ELISA values measured after successful surgical therapy in 2 dogs showed a decrease of anti-P insidiosum antibody concentrations into the normal range as early as 2 months after treatment. We conclude that the ELISA is a sensitive and specific test for the diagnosis of canine pythiosis, and may be a useful tool for monitoring response to medical or surgical therapy.     

Correction to: Molecular analysis indicates high levels of carabid weed seed consumption in cereal fields across Central Europe

Journal of Pest Science -     

Treatment of Intraoperative Persistent Penile Erection in a Stallion

This case report describes the occurrence of persistent penile erection in a breeding stallion that occurred while the horse was under inhalant anesthesia for a carpal arthroscopy. The horse had no history of breeding problems, and no abnormalities were detected on physical examination, complete blood count, or serum chemistry tests performed prior to surgery. Anesthesia was induced with guaifenesin and ketamine after sedation with xylazine and was maintained with isoflurane in 100% oxygen. Penile erection developed approximately 35 minutes after induction and persisted for over 2 hours despite various physical and pharmacological attempts to alleviate it (massage, cold compresses, intravenous benztropine administration, and intracavernosal phenylephrine). Successful resolution of the erection was obtained by cannulation and drainage of blood from the corpus cavernosum and subsequent irrigation with heparinized sterile saline and infusion of phenylephrine in the corpus cavernosum. The detumescent penis was placed back into the sheath, and purse string sutures were placed in the sheath to ensure the penis would remain inside the sheath during recovery. The stallion's recovery from anesthesia was uneventful, the sutures were removed, and the horse was fitted with a penile sling to prevent additional edema or trauma. The stallion recovered completely from the persistent penile erection. Semen was collected 6 days after the event, and he returned to normal pasture breeding 6 weeks after surgery.     

Ammonium‐driven nitrification plays a key role in increasing Mn availability in calcareous soils

Background : Manganese deficiency often becomes a yield limiting factor, particularly on calcareous soils, even though the total soil manganese content is usually sufficient. Although it is known that acidifying N fertilizers can improve Mn availability, the reason of this effect is still unknown. Aim : Our aim was to investigate the effect of stabilized ammonium fertilizers as a tool to distinguish between physiological‐ and nitrification‐induced acidification. Method : Two pot experiments with Triticum aestivum L. and one soil incubation experiment using different nitrogen forms (CN = calcium nitrate, AN = ammonium nitrate, AS = ammonium sulfate, ATS = ammonium thiosulfate) with and without addition of nitrification inhibitors (DCD, Nitrapyrin, Piadin, DMPP) were conducted to examine the effect on Mn availability in the soil and Mn uptake by the plants at different development stages (EC 31 und 39). Results : With increasing fertilizer ${\mathrm{NH}}_4^{+}$ content a higher Mn concentration was detected: CN: 32 µg Mn g^?1 DM, AN: 39 µg Mn g^?1 DW, AS: 55 µg Mn g^?1 DM, ATS: 109 µg Mn g^?1 DM. The addition of a nitrification inhibitor resulted in a significantly lower rhizosphere pH compared to the non‐stabilized fertilizer. Surprisingly, the use of different nitrification inhibitors led to unchanged (CN, AN) or lower Mn concentrations of wheat. Especially in the ${\mathrm{NH}}_4^{+}$ treatments (AS and ATS), this negative effect was very evident (AS+DCD: 42 µg Mn g^?1 DM; ATS+DCD: 55 µg Mn g^?1 DM). Conclusions : Mn availability was enhanced by ongoing nitrification process rather than physiological acidification. Compared to other N forms, ammonium thiosulfate led to the highest Mn availability in bulk soil.     

Comparative study of ovatifolin antioxidant and growth inhibition activities

A comparative study on the effect of arturin (1), ovatifolin (3), deacetylovatifolin (5), and their 1-acetoxyarturin (2), 8-acetoxyovatifolin (4), 1,10-epoxyovatifolin (6), and 11,13-dihydroovatifolin (7) derivatives, isolated from Podanthus ovatifolius and Podanthus mitiqui, on the seedling growth, germination, and respiration of several monocot and dicot weedy target species was carried out. In addition to the inhibitory activity on the bleaching of crocin induced by alkoxyl radicals, these compounds also displayed scavenging properties toward DPPH in TLC autographic and spectrophotometric assays. The results indicate that ovatifolin (3), deacetylovatifolin (5), epoxyovatifolin (6), dihydroovatifolin (7), and the CH(2)Cl(2) extract interfere with pre-emergence of seedlings at the level of respiration. These compounds appear to have selective effects on the radicle and shoot growth of Physalis ixocarpa and Trifolium pratense. Their allelopathic effects are comparable to those of parthenolide, a know natural growth inhibitor.