Deamino-oxytocin and 1-gamma-mercaptobutyric acid-oxytocin: x-ray crystallographic data

X-ray studies of crystalline deamino-oxytocin (1-beta-mercaptopropionic acid-oxytocin) have established the existence of both wet and dry forms. A remarkable degree of similarity exists between the two forms even though there are changes in cell dimensions and space group on drying. The dry form of 1-gamma-mercaptobutyric acid-oxytocin is very similar to that of deamino-oxytocin.     

High Embryonic Recovery Rates with In vivo and Ex vivo Techniques in the Bitch

The embryonic collection techniques in dogs present a vast methodological variation and low recovery rates. The objectives were to compare and describe two techniques as to the recovery of canine embryos, on the 12th day after the first mating or artificial insemination. Embryos were recovered through uterine horn flushing in vivo, before performing the ovariohysterectomy (OHE) (Group 1; n = 9) or ex vivo, immediately after the OHE (Group 2; n = 9). In total, 43 and 47 embryonic structures were recovered in Groups 1 and 2, respectively. There was no significant difference (p > 0.05) between groups on recovery rates (72.8% and 81.0%, respectively). We inferred that both in vivo and ex vivo techniques allow a high rate of embryonic recovery; in the collection technique prior to the OHE, it is essential to carefully handle the reproductive system during the trans‐surgical period and that the 12th day (D12) after the first mating/artificial insemination is an efficient option for the high recovery rate of morulae and blastocysts.     

Restricted lateral diffusion of PH-20, a PI-anchored sperm membrane protein

The rate of lateral diffusion of integral membrane proteins is constrained in cells, but the constraining factors for most membrane proteins have not been defined. PH-20, a sperm surface protein involved in sperm-egg adhesion, was shown to be anchored in the plasma membrane by attachment to the lipid phosphatidylinositol and to have a diffusion rate that is highly restricted on testicular sperm, being more than a thousand times slower than lipid diffusion. These results support the hypothesis that lateral mobility of a membrane protein can be regulated exclusively by interactions of its ectodomain.     

Flowering behaviour and seed fertility in dihaploidSolanum tuberosum

Summary Aspects of female fertility were studied in clones from the Pentlandfield dihapoid collection which has been derived predominantly from British tetraploid potatoes. Dihaploids from different tetraploid parents differed significantly in such features as flower initiation and bud abscission. Compared with tetraploids, a higher proportion of dihaploid clones reached anthesis but otherwise their flowering behaviour was similar. They produced fewer ovules per ovary and set less seed which was of lower viability. Taken together the results indicated a considerable reduction in female fertility in the first dihaploid generation.

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Zusammenfassung Zur Untersuchung der Dihaploiden der 1. Generation dienten zum Teil Klone aus der Pentlandfield-Dihaploidkollektion, die meistens aus britischen Züchtungen und aus Züchtungsmaterial von Elite-Tuberosum stammen. Bei 181 Dihaploiden von 20 verschiedenen tetraploiden Eltern wurden Aspekte des Blühverhaltens notiert (Tabelle 1). Statistisch gesicherte Unterschiede wurden zwischen Elterngruppen im Anteil der Klone die Blüten hervorbrachten, und im Anteil der Klone, die alle Knospen abwarfen, gefunden. Insgesamt war der Prozentsatz an Dihaploiden, die zum Erblühen kamen (40,9%), signifikant h?her als in einem grossen Muster von Tetraploiden (28,2%; East, 1908), w?hrend der Prozentsatz jener, die alle Knospen abwarfen, geringer war (23,7% gegen 70,6%). Dies kann mit der unvermeidlichen Auslese von mehr freiblühenden tetraploiden Klonen für die Erzeugung von Dihaploiden im Zusammenhang stehen. Wenn wir jedoch den durchschnittlichen Verlust von offenen Blüten durch Abwurf (13,7%) mit jenem von 16 Tetraploiden (11,6%) vergleichen, so war der Unterschied klein. Dihaploide Klone, deren Blühf?higkeit von Jahr zu Jahr schwankte, brachten gew?hnlich wenig Blüten, und ihr Verhalten wurde wahrscheinlich sowohl von genetischen als auch von Umweltfaktoren bestimmt (Tabelle 2); darin glichen sie dem tetraploiden Tuberosum (J?hnl, 1954). Die Anzahl der Samenanlagen in den Fruchtknoten von dihaploiden und tetraploiden Eltern wurde durch Zerlegung und Ausz?hlung unter einem Mikroskop mit schwacher Vergr?sserung (Tabelle 3) bestimmt. Ein statistischer Vergleich zwischen Dihaploiden und einem Muster von Tetraploiden ergab hochsignifikante Unterschiede in der Anzahl Samenanlagen, sowohl zwischen Klonen innerhalb jeden Ploidiegrades als auch zwischen den Ploidiegraden: dihaploider Durchschnitt =483,9 pro Fruchtknoten; tetraploider Durchschnitt =732,3. Dies würde dazu führen, die Fruchtbarkeit der Dihaploiden herabzusetzen, indem die Anzahl der Samen, die durch Best?ubung erzielt werden, beschr?nkt wird. Unterschiede zwischen dihaploiden Klonen in der Anzahl unbefruchteter Samen sind nicht auf die tetraploide Elternschaft zurückzuführen. Der Prozentsatz an Samenanlagen, die nach der Testkreuzung als Samen gewonnen wurden, war bei Dihaploiden viel kleiner als bei Tetraploiden, so dass der fertilste Dihaploid nur 10,3% gegenüber 19,7% des am wenigsten fertilen Tetraploids erbrachte. Die Daten über die Samenfruchtbarkeit wurden bei 50 normal blühenden Dihaploiden in Testkreuzungen mit diploiden Phureja-Klonen von geprüfter m?nnlicher Fertilit?t erlangt (Tabelle 4). Die Dihaploiden unterschieden sich signifikant in den Anteilen an Testbest?ubungen, die Beeren erbrachten, aber es bestand keine nachweisbare Beziehung zwischen Beerenansatz und Anzahl Samen pro Beere. 25 der 50 Klone brachten keinen Samenansatz, ein gr?sserer Anteil als von den blühenden Tetraploiden erwartet. Der Durchschnitt von 5,2 Samen pro Best?ubung für Dihaploide war weniger als ein Drittel wie bei Dihaploid × Phureja Hybriden (Carroll, im Druck), wenn man annimmt, dass die festgestellte Depression der weiblichen Fertilit?t bei den Dihaploiden auf ihre Inzuchtnatur (relativ) zurückzuführen ist. Die Angaben über die Samenkeimung (Tabelle 4) zeigten in diesem Stadium ebenfalls eine reduzierte Lebensf?higkeit der Samen.

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Résumé La première génération de dihaplo?des étudiée forme une partie de la collection dihaplo?de de Pentlandfield de clones dérivés principalement de cultivars britanniques et de matériel élite d’amélioration Tuberosum. Les aspects de la floraison ont été enregistrés sur 181 dihaplo?des provenant de 20 parents tétraplo?des différents (tableau 1). On trouve des différences statistiquement significatives entre les groupes parentaux dans la proportion de clones commen?ant à fleurir, et la proportion subissant l’abscission de tous les bourgeons. Au surplus, le pourcentage de dihaplo?des atteignant l’anthèse (40,9%) est significativement plus élevé que dans un vaste échantillon de tétraplo?des (28,2%; East, 1908), tandis que le pourcentage perdant tous les bourgeons était plus bas (23,7%, contre 70,6%). Ceci peut être lié à l’inévitable sélection de clones tétraplo?des fleurissant plus librement pour la production dihaplo?de. Cependant, quand nous comparons la perte moyenne par abscission (13,7%) de fleurs ouvertes avec celle de 16 tétraplo?des (11,6%), il y a peu de différence. Les clones dihaplo?des dont la capacité de fleurie varie de saison à saison habituellement sont faiblement florifères et leur comportement est probablement déterminé par la génétique aussi bien que par les facteurs environnants (tableau 2); en ceci ils ressemblent au Tuberosum tétraplo?de (J?hnl, 1954). Les nombres d’ovules dans les ovaires de parents dihaplo?des et tétraplo?des sont déterminés par dissection et comptage sous microscope à faible puissance. (tableau 3). Une comparaison statistique entre dihaplo?des et un échantillon de tétraplo?des donne des différences fautement significatives dans les nombres d’ovules, à la fois entre clones dans chaque niveau de plo?die et entre niveaux de plo?die: moyenne dihaplo?de =483,9 par ovaire; moyenne tétraplo?de: 732,3. Ceci tend à réduire la fertilité dihaplo?de en diminuant le nombre de semences que l’on peut obtenir par pollination. Les différences entre clones dihaplo?des dans les nombres d’ovules ne sont pas en relation avec le parentage tétraplo?de. Le pourcentage d’ovules donnant des semences après croisement de test est beaucoup plus bas chez les dihaplo?des que chez les tétraplo?des, à tel point que le dihaplo?de le plus fertile donne seulement 10,3% contre 19,7% dans le tétraplo?de le moins fertile (tableau 3). Les données sur la fertilité des semences proviennent de 50 dihaplo?des fleurissant normalement utilisés dans des croisements-tests avec des clones diplo?des Phureja de fertilité male démontrée (tableau 4). Les dihaplo?des différent significativement dans les proportions de pollinations-tests qui donnent des baies mais il n’y a aucune relation démontrable entre l’assortiment de baies et le nombre de semences par baie. Vingtcinq des 50 clones ne produisaient pas de semences, une proportion plus élevée que l’on pourrait atteindre avec des tétraplo?des fleurissant. La moyenne de 5,2 semences par pollination pour les dihaplo?des est inférieure d’un tiers à celle des dihaplo?des × Phureja hybrides (Carroll, sous presse), ce qui fait supposer que la dépression marquée de la fertilité femelle dans les dihaplo?des peut être attribuée à sa nature relativement autogame. Les données sur la germination des semences (tableau 4) montrent une viabilité réduite à ce stade également.

     

Production of Oocytes of Nile Tilapia (Oreochromis niloticus) for In Vitro Fertilization via Hormonal Treatments

Only a few studies have described hormonal treatments for induction of synchronicity and gamete collection in Nile tilapia (Oreochromis niloticus), both important for assortative matings in breeding programmes and essential for polyploidy technologies. In this study, we compared the effectiveness of carp pituitary extract (CPE), Nile tilapia pituitary extract (TPE), human chorionic gonadotropin (hCG) and gonadotropin‐releasing hormone (GnRH) protocols on the induction of spawning and egg production in Nile tilapia. Among the hormonal treatments analysed, only hCG was effective for producing viable gametes for in vitro fertilization. To verify the viability of this hormonal treatment, hCG was tested using different doses (1000, 2000, 3000, 4000 and 5000 IU/kg) in a large number of females (208 animals) from two Nile tilapia lines. The results indicated that hCG doses between 1000 and 5000 IU/kg could be used to induce final oocyte maturation in Nile tilapia with collection of stripped oocytes. This is the first study to report differential reproductive responses to hormonal treatment between tilapia lines: line 1 was more efficient at producing eggs and post‐hatching larvae after hCG induction than line 2. In conclusion, we demonstrated that the hCG protocol may be applied on a large scale to induce final oocyte maturation in Nile tilapia. The development of a protocol for in vitro fertilization in Nile tilapia may aid in breeding programmes and biotechnological assays for the development of genetically modified lines of Nile tilapia.     

Study of Sperm Concentration,Seminal Plasma Composition and their Physiological Correlation in the Persian Sturgeon,Acipenser persicus

The objectives of the present study were to determine ionic and organic composition of seminal plasma, sperm concentration and their relationships in the Persian sturgeon (Acipenser persicus). In this regard, ionic content (Na⁺, K⁺, Cl^?, Ca²⁺ and Mg²⁺) and organic content (total protein, glucose, cholesterol and triglyceride) along with sperm concentration were measured in 17 specimens of the Persian sturgeon. The seminal plasma contained 59.53 ± 2.56 mm /l sodium, 9.1 ± 1.42 mm chloride, 4.72 ± 0.3 mm potassium, 1.45 ± 0.075 mm calcium and 0.7 ± 0.072 mm magnesium. The following organic contents were found: total protein 0.11 ± 0.02 g/dl, glucose 22.18 ± 4.16 mg/dl, cholesterol 6.67 ± 1.04 mg/dl and triglyceride 15.2 ± 0.65 mg/dl. The mean sperm concentration was estimated to be 1.6 ± 0.12 (×10⁹ sperm/ml). A significant relationship was found between sperm concentration and K⁺ of seminal plasma (r = 0.533, p < 0.05). Significant correlations were observed between ionic contents: Na⁺ vs Cl^? (r = ?0.854, p < 0.01) and Mg²⁺ vs K⁺ (?0.583, p < 0.05). Also, level of triglyceride was negatively correlated with Mg²⁺ (r = ?0.503, p < 0.05). Presented data could be considered as a complementary study for developing special extenders and protectant solutions for improving artificial fertilization in this valuable species.     

Infectivity and persistence of an outbreak strain of Salmonella enterica serotype Typhimurium DT160 for house sparrows (Passer domesticus) in New Zealand

AIM: To examine the infective dose, incubation period and disease progression of an isolate of Salmonella enterica serotype Typhimurium definitive type 160 (DT160) originating from a naturally-infected house sparrow (Passer domesticus) during an outbreak of the disease in New Zealand.

METHODS: Thirty-six house sparrows captured from the wild and free of Salmonella spp were divided into six groups of six birds, housed individually, and inoculated orally with phosphate buffered saline (PBS) or 10¹, 10², 10³, 10⁵, 2 × 10⁸ colony forming units (cfu) of the outbreak strain of S. Typhimurium DT160. The birds were observed for 10 days for clinical signs and/or mortality, and faecal samples were collected to determine excretion of S. Typhimurium. The birds were eutha- nised 11 days post-inoculation (p.i.) and a wide range of tissue samples were collected for histopathological examination, and culture and typing of Salmonella spp. Macro-restriction profiling by pulsed-field gel electrophoresis (PFGE) using XbaI was performed for the epidemiological typing of S. Typhimurium DT160 isolates.

RESULTS: Mortality in house sparrows inoculated with S. Typhimurium DT160 was dose-dependent, and 2/6 birds inoculated with ¹⁰⁵ cfu and all six birds inoculated with 2 × 10⁸ cfu died during the study. Infected sparrows displayed few clinical signs, apart from diarrhoea and/or polyuria, fluffed plumage, and sitting on the floor of the cage. Faecal excretion of DT160 occurred briefly in two birds inoculated with 10² cfu and four birds inoculated with 10³ cfu, on most days in five birds inoculated with 10⁵ cfu, and continuously in six birds inoculated with 2 × 10⁸ cfu. DT160 was isolated from the livers of three birds which received 10³ cfu, five birds dosed with 10⁵ cfu, and all six birds given 2 × 10⁸ cfu. Following necropsy, histopathological lesions similar to those seen in the natural disease were observed in the liver or spleen of three birds which received 10³ cfu, and all birds dosed with ≥10⁵ cfu.

CONCLUSION: The results indicate that an isolate of S. Typhimurium DT60 originating from house sparrows in New Zealand is pathogenic to these birds and that the response is dose- dependent. The persistence and excretion of the pathogen may last for at least 10 days. This confirms that sparrows infected with DT160 could be a source of infection to humans and other in-contact animals.     

Crypt abscesses in the caeca of feral rabbits in a rabbit haemorrhagic disease endemic region of New Zealand

AIM: To describe the pathology of crypt abscesses in the caeca of feral rabbits in the Manawatu region of New Zealand, and to examine the possible relationship between their prevalence and rabbit haemorrhagic disease (RHD) virus (RHDV) infection.

METHODS: During the course of a 3-year study of RHD, 173 wild rabbits (Oryctolagus cuniculus) were shot on three pastoral livestock farms in the Manawatu region of New Zealand. All rabbits were necropsied, and tissue samples of the caeca were examined histologically. The age of each animal was determined, and blood samples collected for the detection of RHDV antibodies. Logistic regression was used to model the odds of rabbits having crypt abscesses.

RESULTS: At necropsy, 63/173 (36.4%) rabbits were found to have small circular black nodules on the mucosal surface of their appendix caecum and/or sacculus rotundus. Microscopically, these were identified as small crypt abscesses composed of dilated sacs at the base of the mucosa that were often lined by a thin layer of attenuated epithelial cells. They usually contained large amounts of concentrically-laminated mucopolysaccharide material that was sometimes pigmented, inflammatory debris, and were often the site of a moderate multifocal appendicitis. Although RHDV was active in the study area, no association was found between RHDV antibodies in serum and the presence of lesions. The lesions were more common in older individuals and those born in summer or autumn.

CONCLUSIONS: This is the first report of crypt abscesses with inflammation and necrotic debris in the caeca of rabbits. No association between the occurrence of crypt abscesses and RHDV infection was identified. Wild rabbits born in a particular season were presumably exposed very early in life to conditions that caused the crypt abscesses to develop. Alternatively, association with season of birth may represent rabbits that were of similar age in a later stage of their lives, when they became exposed to the cause of the lesions, which remains unidentified.     

Concurrent avian malaria and avipox virus infection in translocated South Island saddlebacks (Philesturnus carunculatus carunculatus)

Abstract

CASE HISTORY: Outbreaks of mortality in South Island saddlebacks (Philesturnus carunculatus carunculatus) that had been translocated to two offshore islands in the Marlborough Sounds of New Zealand were investigated during the summer of 2002 and 2007. Both outbreaks were associated with a severe decrease in numbers of saddlebacks of up to 60% of approximately 200 birds.

CLINICAL AND PATHOLOGICAL FINDINGS: Many of the surviving birds were in poor condition, and had skin lesions on the legs and head. Necropsy showed pale liver and lungs, and a swollen spleen. Histopathology revealed schizonts resembling Plasmodium spp. within the cytoplasm of many hepatocytes and splenic histiocytes. The skin lesions consisted of epithelial proliferations containing numerous Bollinger bodies typical of avipox virus (APV) infection. Two different APV were isolated, using PCR, from two different birds exhibiting skin lesions. Each isolate had 100% sequence homology with APV members from either Clade A or Clade B. In addition, PCR analysis revealed that the Plasmodium elongatum present in infected birdsbelonged to a strain that was endemic in the population of North Island saddlebacks (Philesturnus carunculatus rufusater).

DIAGNOSIS: Concurrent infections with Plasmodium spp. haemoparasites and APV were identified as the likely cause of death in the birds examined.

CONCLUSIONS AND CLINICAL RELEVANCE: Although the Plasmodium spp. identified is thought to be endemic to saddlebacks in New Zealand, the affected birds were likely to be immunocompromised by concurrent APV infection or through lack of genetic diversity. Both the introduced mosquito Culex quinquefasicatus and the native mosquito Culex pervigilans are likely vectors for both these diseases, and the provision of water supplies less favourable to mosquito-breeding is recommended.