A rapid monoclonal immunofluorescence assay for Chlamydia psittaci in fecal smears from psittacine birds

One hundred two fecal specimens from psittacine birds submitted to Veterinary Laboratory Services of the California Department of Food and Agriculture at Petaluma were screened for Chlamydia psittaci by a direct immunofluorescence assay using a fluorescein-labeled monoclonal antibody conjugate specific for Chlamydia sp. Results were compared with those obtained by isolation of chlamydia in cultures of McCoy mouse cells. The relative specificity of the direct fluorescent antibody test on fecal smears was 98.9% and the relative sensitivity was 62.5%. The results of this study suggested that the direct fluorescent antibody test was highly specific, and it proved to be a useful same-day antemortem diagnostic test for birds with symptomatic chlamydial infection. The use of centrifugation in the cell culture assay was found to significantly enhance the level of chlamydial infection in cell culture.     

Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.     

Feed preference and growth response of juvenile Litopenaeus vannamei to supplementation of marine chemoattractants in a fishmeal‐challenged diet

This study compared the feed preference and growth response of Litopenaeus vannamei to chemoattractants. A diet with 3% fishmeal was supplemented with either 3% salmon meal (POS), 3% soy protein concentrate (NEG), 3% krill meal (KRM), 3% squid meal (SQM), 3% shrimp head meal (SHM), 3% shrimp meal (SM), 3% squid liver meal (SLM), or 5% liquid sardine hydrolysate (SAH). Shrimp with a body weight (BW) of 0.99 ± 0.08 g were stocked at 100 animals/m² in 56 tanks of 1 m³ and fed 10 times daily for 74 days. Feed preference was evaluated by feeding shrimp of 10.87 ± 1.82 g in excess twice a day for 10 days in two separate feeding trays allocated in 50 tanks of 0.5 m³. Survival reached 93.3 ± 5.80% and was unaffected by supplementation. Final BW was the highest for shrimp fed the KRM‐supplemented diet (11.97 ± 0.93 g), followed by POS (11.11 ± 0.77 g) and SQM (11.01 ± 1.17 g). Diets SHM, SM, SLM, and NEG showed a lower shrimp BW than POS, but were not statistically different among them. Shrimp fed the SAH diet achieved the lowest BW (10.06 ± 1.02 g). The highest gained yield was obtained with diets KRM and POS. No statistical difference was observed in shrimp yield among other diets. The lowest feed conversion ratio (FCR) was achieved with shrimp fed KRM (1.31 ± 0.05) when compared to diets SHM (1.47 ± 0.05), SAH (1.47 ± 0.07), and SLM (1.45 ± 0.17). Two‐by‐two comparisons indicated that shrimp preferred SHM and KRM, except when these were compared to SQM and SLM. No difference in feed preference was found between diets with SQM and SLM. SAH was the least preferred raw material in all comparisons. Results indicated that KRM acts as a powerful feeding effector and growth enhancer in fishmeal‐challenged diets for whiteleg shrimp. A dietary supplementation with 3% KRM is more effective than the same dose of any other chemoattractant evaluated.     

Cultivation of Pen Shells: An Example with Atrina maura in Northwestern Mexico

Pen shells are economically important and most are threatened by overexploitation and habitat degradation. Cultivation represents an alternative for preserving the resource and increasing the production. Atrina maura was cultivated in a suspension system which was followed by a bottom phase. Samples of pen shells and tissues were collected monthly to measure absolute growth, shell height increase, and an adductor muscle index. After the trials, adults shell height was 195 ± 10 mm, total weight was 223 ± 28 g, and mean shell height increase was 0.81 cm/mo. Protective devices were vital to avoid predation by crabs and fish; survival was 90% in the suspension phase and 70% in the bottom phase. Variations in the muscle index were apparently related to reproduction, suggesting activity (partial spawning) from August 2008 to April 2009 in juveniles and a major spawning in adults during August 2009. Adductor muscles reached a mean of 21 g after 20 mo of cultivation, but muscle weights decreased significantly after reproduction (August to November 2009). Results indicated that A. maura is suitable for aquaculture and the cultivation system adopted here should be tested in pilot‐scale ventures. This system may work with other species of pen shells.     

Flash flood simulation and valve behavior of Mytilus galloprovincialis measured with Hall sensors

Mussels close their shell as a protective strategy and the quantification of this behavioral marker may represent an alarm signal when they are exposed to environmental stressors. In the present study, we investigated the ability of the Mediterranean mussel Mytilus galloprovincialis to recover and then the resilience or inertia of valve activity after a pulsing exposition to diverse levels of salinity (5, 10, 20, and 35 PSU as reference value). The trial simulated an event of drastic and sudden reduction of seawater salinity thus mimicking an event of flash flood from intense rain. Valve gaping and movements were measured in continuous cycle for 10 days using a customized magneto-electric device which uses Hall sensors. Results showed that under normal conditions of salinity (35 PSU), the general pattern of valve movements was a continuously open state with sporadic spikes indicating a closing motion. At salinity of 5, PSU mussels reacted by closing their valves, leading to a 77% mortality on the 4th day. At salinity of 10, PSU animals were observed with closed valves for the entire duration of the exposure and no mortality occurred, they showed a significant reduction in the valve activity once the reference value of salinity was re-established. In contrast, salinity of 20 PSU did not trigger a significant behavioral response. Interestingly, there no define rhythms of valve movements were recorded during salinity challenges.     

Canine squamous cell carcinoma: Electrochemotherapy association with surgery and correlation with overall survival

Squamous cell carcinoma (SCC) is an important malignancy in dogs, due to its incidence and clinical presentation, which can be of locally aggressive single or multiple lesions with a metastatic potential. The objective of this investigation was to evaluate SCC response to treatment, anatomopathological and immunohistochemical characteristics, disease-free interval and overall survival time. 54 dogs with histopathologically diagnosed SCC were included in this study. Their mean age was 9.16 years with a range of 1–14 years. Of the 54 animals in the study, 34 (65.4%) had white skin and white fur coats. There was a significant correlation between fur coat colour and the development of tumours in areas of sun exposure (p = .001). Animals with tumours in areas of the body exposed to the sun had longer overall survival time than animals with tumours in areas not associated with sun exposure (p = .001). Surgery combined with electrochemotherapy (ECT) yielded a survival rate 32% higher than using a surgical approach alone (HR = 0.32, p = .038, IC = 0.11–0.94). ECT, with or without surgery, had an objective response rate of 90.9%. Local lymph node and/or distant site metastasis at diagnosis, or at some point during follow-up, occurred in 34.6% (18/52) of animals. Animals with tumours in sun exposed locations had more aggressive histopathological characteristics but had longer overall survival time. This is probably due to individualised therapeutic treatment with both surgery and ECT.     

Verrucosamide,a Cytotoxic 1,4-Thiazepane-Containing Thiodepsipeptide from a Marine-Derived Actinomycete

A new cytotoxic thiodepsipeptide, verrucosamide (1), was isolated along with the known, related cyclic peptide thiocoraline, from the extract of a marine-derived actinomycete, a Verrucosispora sp., our strain CNX-026. The new peptide, which is composed of two rare seven-membered 1,4-thiazepane rings, was elucidated by a combination of spectral methods and the absolute configuration was determined by a single X-ray diffraction study. Verrucosamide (1) showed moderate cytotoxicity and selectivity in the NCI 60 cell line bioassay. The most susceptible cell lines were MDA-MB-468 breast carcinoma with an LD₅₀ of 1.26 µM, and COLO 205 colon adenocarcinoma with an LD₅₀ of 1.4 µM. Also isolated along with verrucosamide were three small 3-hydroxy(alkoxy)-quinaldic acid derivatives that appear to be products of the same biosynthetic pathway.     

First isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata (Moreau)

The first isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata, is reported. The pathogen was recovered from ulcers of cultured fish, from three different outbreaks. The six isolates obtained were biochemically and serologically characterized and diagnosis was confirmed by polymerase chain reaction using specific primers and partial 16S rRNA gene sequencing. The isolates constituted a homogeneous phenotypic group; however, they belong to two of the different serotypes described within this species. A virulence evaluation of the isolates using Wedge sole fry was also performed.