Stickstoff-Assimilation in der Wurzel und Transport von N-Verbindungen mit dem Blutungssaft der Wurzeln in Abhängigkeit von der Mn-Versorgung

Nitrogen Assimilation in Roots and the Transport of Nitrogen Compounds in the Bleeding Sap of Roots in relation to Manganese Nutrition. The assimilation of nitrogen in the roots of 27 days old pumpkin plants was examined in relation to manganese nutrition. The transport of nitrogen compounds in the xylem was determined in roots and in the bleeding sap of roots using nitrate as the N-source. The maximum NO₃ content in the roots was observed in the Mn treatment which resulted in the highest shoot yields (0.05 ppm Mn). The bleeding sap of this treatment was lowest in nitrate concentration, but showed the highest rate of transport of organic nitrogen compounds. In experiments with ¹⁵N in the nutrient solution the isotope was found in the roots in organic and in inorganic compounds. The composition of the fraction of free amino acids differed between roots and xylem sap. In the bleeding sap glutamine was especially dominant. In the roots the amino acid composition depended on the extent of Mn-supply. Lowest glutamine concentrations were found in the xylem sap from the treatment with maximum shoot yields. A numerical difference was found in the xylem sap between organic N (N_(org)) and the amino acid nitrogen. This difference which account for more than 50 % of the organically bound nitrogen is suggested to be made up in part by low molecular weight peptides, amino sugars and other compounds. In Mn deficiency a general reduction in the intensity of nitrogen metabolism was found. With Mn toxicity the N assimilation activity was more intensive than for the low Mn supply. Simultaneously, however, the transport of organic N compounds from the root was lower.     

In Vitro Maturation and Fertilization of Buffalo Oocytes: Effects of Storage of Ovaries, IVM Temperatures, Storage of Processed Sperm and Fertilization Media

Studies were conducted to examine the possibility of preserving slaughterhouse‐derived buffalo ovaries at 4°C for 0 (control), 12 and 24 h to maintain the developmental competence of the oocytes (experiment 1), to assess the effect of incubation temperature during oocyte maturation on rates of in vitro maturation (IVM) and in vitro fertilization (IVF) of buffalo oocytes and embryo development (experiment 2), and to examine the effect of storage at 25°C for 0 (control), 4 and 8 h of frozen–thawed buffalo sperm and BO and H‐TALP as sperm processing and fertilization media on cleavage and embryo development in vitro of buffalo oocytes (experiment 3) in order to optimize the IVF technology in buffalo. Results suggested that storage of ovaries at 4°C for 12 or 24 h significantly (p < 0.05) reduced the developmental potential of oocytes. Incubation temperatures during the IVM influenced the fertilization rate but had no significant effect on maturation and subsequent embryo development. The incubation temperature of 38.5°C during IVM was found to be optimum for embryo production in vitro. Storage of frozen–thawed sperm at 25°C for 8 h significantly (p < 0.05) decreased its ability to cleave the oocytes. Sperm processed in BO medium had significantly (p < 0.05) higher ability to cleave the oocytes than the H‐TALP medium.     

Antimicrobial susceptibility of bacteria isolated from neonatal foal samples submitted to a New Zealand veterinary pathology laboratory (2004 to 2013)

AIMS: To describe antimicrobial susceptibility, and identify antimicrobial resistance (AMR), in bacteria isolated from New Zealand foals.

METHODS: A database search was performed of submissions to a veterinary pathology laboratory between April 2004 and December 2013 for bacterial culture of samples from foals <3 weeks of age. Culture and susceptibility results were compiled with demographic information. Susceptibility results were as defined for the Kirby-Bauer disk diffusion susceptibility test based on Clinical Laboratory Standards Institute guidelines. Multi-drug resistance (MDR) was defined as non-susceptibility to ≥3 of a panel of antimicrobials (ceftiofur, enrofloxaxin, gentamicin, penicillin, tetracycline, trimethoprim-sulfonamide); penicillin susceptibility was not included for Gram-negative isolates.

RESULTS: Submissions from 102 foals were examined, and 127 bacterial isolates were cultured from 64 (63%) foals. Of the 127 isolates, 32 (25%) were Streptococcus spp., 30 (24%) were Staphylococcus spp., 12 (10%) were Enterococcus spp. and 26 (21%) were Escherichia coli. Of 83 Gram-positive isolates, 57 (69%) were susceptible to penicillin. Over all isolates, 92/126 (73%) were susceptible to gentamicin and 117/126 (93%) to enrofloxacin; 62/82 (76%) of Gram-positive, and 22/42 (52%) of Gram-negative bacteria were susceptible to ceftiofur; 53/81 (65%) of Gram-positive, and 23/44 (52%) of Gram-negative bacteria were susceptible to tetracycline; 59/82 (72%) of Gram-positive, and 23/44 (43%) of Gram-negative bacteria were susceptible to trimethoprim-sulfonamide. Of 126 isolates, 33 (26%) had MDR; >1 isolate with MDR was cultured from 24/64 (38%) foals, and ≥2 isolates with MDR were recovered from 8/64 (13%) foals.

CONCLUSIONS: Multi-drug resistance, including resistance to commonly used antimicrobials, was found in bacterial isolates from foals in New Zealand.

CLINICAL RELEVANCE: The results of this study are of concern from a treatment perspective as they indicate a potential for antimicrobial treatment failure. For future surveillance of AMR and the creation of national guidelines, it is important to record more data on samples submitted for bacterial culture.     

Comparison of Whole Ovary Cryotreatments for Fertility Preservation

The goal of this study was to compare a traditional slow‐freeze method (TF) with an open unidirectional slow freeze cooling system (UF) for whole ovary cryopreservation. Therefore, whole pig ovaries were randomly assigned to (A) fresh control, (B) traditional slow freeze (TF) or (C) unidirectional slow freeze (UF). Ovaries were perfused with 10% DMSO in Krebs‐Ringer. For TF, whole ovaries were placed in specimen jars containing 10% DMSO and placed into a specialized container for freezing filled with propan‐2‐ol. For UF, whole ovaries were placed within a specially designed container containing 10% DMSO and transferred to a specialized freezing machine (CTE 920). Histological evaluation demonstrated intact morphology of follicles in all groups; however, an overall decrease of follicle numbers in TF (46%) and UF (50%) compared to fresh control. Live/dead assay indicated significantly lower populations of live cells in both TF (60%) and UF (58%) compared to fresh tissue (74%). TUNEL assay confirmed a difference in percentage of apoptotic follicles between fresh and TF, but there was no significant difference between fresh and UF. To improve the structural and functional integrity of whole ovaries, further investigation, especially into directional freezing, is needed. Whole ovary cryopreservation could provide opportunities for women facing fertility loss due to chemo‐ or radiotherapy treatment.     

Elevated temperature affects growth and hemato‐biochemical parameters,inducing morphological abnormalities of erythrocytes in Nile tilapia Oreochromis niloticus

Temperature is considered as an important environmental factor, and the increasing water temperature resulting from global warming is a great concern. The present study was conducted to examine the effects of elevated water temperature on growth, hemato‐biochemical parameters in Nile tilapia, Oreochromis niloticus acclimatized to three temperatures (31°C, 34°C and 37°C) for 60 days. Additionally, erythrocytic cellular abnormalities (ECA) and erythrocytic nuclear abnormalities (ENA) tests were assayed using peripheral erythrocytes after exposure to the three temperatures. Fish were sacrificed on days 7, 15, 30 and 60 of exposure. Growth performances viz., weight gain, % weight gain and specific growth rate (SGR) showed decreasing tendency at 34°C but significantly declined at 37°C compared to 31°C. The abundance of haemoglobin (Hb) and red blood cells (RBCs) significantly decreased in response to temperature increases, while white blood cells (WBCs) displayed the opposite response. At days 7 and 15, blood glucose levels significantly increased in response to the temperature increase, while at days 30 and 60 glucose declined. Frequencies of ECA and ENA were significantly enhanced at the highest temperature throughout the experimental period. Dissolved oxygen decreased and free CO₂ increased significantly with increasing temperature throughout the study period. The present study revealed that temperatures higher than 34°C may be hazardous to O. niloticus.     

Resident forum abstractsEVALUATION OF THROMBOELASTOGRAPHY (TEG) IN NORMAL CATS

Objective: To establish normal parameters of thromboelastography (TEG) in healthy adult cats. Background: Thromboelastography (TEG) is an in vitro test of coagulation that has been shown to be useful in humans, dogs and select species to identify and quantify alterations of hemostasis (e.g., hypercoagulable and hypocoagulable states). It has also been demonstrated to be useful in monitoring effects of anticoagulant therapies. This test has not been evaluated in cats. Methods: Blood was collected from 25 clinically normal cats by venipuncture using a 21 gauge×3 1/2 inch butterfly catheter and syringe for medial saphenous or jugular venipuncture. A single 1.8 mL sample in 3.8% Sodium Citrate (9:1) was collected from each cat. Recalcified whole blood was analyzed 30 minutes following collection with the TEG® 5000 analyzer (Haemoscope, Niles, IL). Analysis temperature was 37.6°C. TEG parameters recorded included: R‐value (represents initial fibrin formation), K (time from R to standard fixed measure of clot firmness which represents contributions of platelets and fibrinogen), maximum amplitude (MA; represents absolute clot strength), and alpha angle (α; the slope of TEG tracing which represents rate of clot formation). The coagulation index (CI) was derived from the formula generated for humans to provide an overall assessment of whether the sample was hyper‐ or hypocoagulable. Results: Values for the 25 normal cat samples are reported as mean ±2 standard deviations. R=2.97; 1.23–4.72; K=1.54, 0.38–2.71; α=70.70, 57.76–83.65; MA=58.50, 45.26–71.74 and CI=2.27, 0.07–4.46. Compared to historical information obtained on normal dogs, cats have significantly shorter R and K and larger α, MA and CI. Conclusions: TEG does have reproducible performance when used to evaluate coagulation status in normal cats. Compared to dogs, normal cats favor a hypercoagulable state. Species‐specific normal values are necessary for interpretation of TEG results. This test bears potential value for use in future experimental and clinical work to investigate hemostasis in cats receiving anticoagulant therapies or in cats suffering from diseases such as cardiomyopathy which are thought to be associated with altered coagulation status.     

Milk and milk components reduce the motility of Ostertagia circumcincta larvae in vitro

AIM: To examine the effects in vitro of bovine milk and milk products and soymilk on the motility of sheathed and exsheathed L3 Ostertagia circumcincta (also known as Teladorsagia circumcincta) as a measure of larval viability and infectivity. METHODS: L3 were exsheathed in 0.2% sodium hypochlorite, resuspended in Hank's Balanced Salt Solution (HBSS) pH 7.4 and incubated with test solutions at 37 degrees C for up to 48 h. The motility of 50 larvae from each incubate was assessed at selected times using a McMaster slide. Larvae were considered immotile only if straight and not moving. Fresh bovine milk, homogenised milk (3.3% fat), low-fat milk (0.2% fat) and lamb milk replacer were diluted with HBSS pH 7.4 to concentrations from 1.6-100%, and incubated with exsheathed L3 for 1, 24 or 48 h. Bovine whey protein was tested in concentrations of 5-15% at pH 2.5-6.5, casein at 5 or 7.5%, and skim milk powder from 5-15% at pH 5.5 or 6.5, all for 2, 4 or 24 h. Soymilk was tested in concentrations of 1.6-100% for 1, 2, 24 or 48 h. HBSS was used as the control solution. Sheathed L3 were incubated in HBSS pH 7.4, 50% homogenised milk in HBSS, or 50% soymilk in HBSS. Each solution was incubated for 1,2, 24 or 48 h. RESULTS: The motility of exsheathed L3 was reduced by fresh bovine milk, homogenised milk, low-fat milk, lamb milk replacer, whey, casein and skim milk solutions, but not by soymilk. The mean percentage (and SE) immotile at 48 h were: fresh milk 38% (SE 20); homogenised milk 65% (SE 7); low-fat milk 57% (SE 5); lamb milk replacer 43% (SE 7); and soymilk 7% (SE 0.5). Larval immotility increased in whey protein solutions from 5-15%, from pH 2.5-6.5 and from 2 to 24 h (all p<0.001); in skim milk from 5-15% (p<0.001), and was greater at pH 6.5 than at pH 5.5 (p<0.001); in casein from 5-7.5% (p<0.001), but was no different at pH 5.5 and 6.5. The motility of sheathed L3 was reduced at 24 h (p=0.009) and 48 h (p<0.001) by 50% homogenised milk, but not by 50% soymilk or HBSS. CONCLUSIONS: Bovine milk proteins, or components associated with the proteins, reduced the motility of both sheathed and exsheathed L3 O. circumcincta. Soymilk had no effect on nematode motility. Lower larval motility may reduce worm establishment and be a contributing factor to the smaller burdens of gastrointestinal nematodes in milk-fed animals compared with animals after weaning.