Recent developments and improvements in ornamental fish packaging systems for air transport

Current ornamental fish packaging systems are characterized by very high fish loading densities and high metabolic wastes in the transport water after shipment. They focus mainly on management of the quality of transport water. Recent studies using the guppy as a model fish showed that post‐shipment mortality could be reduced through enhancement of the stress resistance of the fish, and hence emphases should also be placed on the preparation of the fish for transport and recovery of the fish after shipment. Farmers can contribute significantly by applying nutritional prophylaxis before harvesting. Exporters may use the salinity stress test to identify fish lots of good quality for transport, apply health prophylaxis to eradicate parasites and optimize other techniques such as starvation of the fish or addition of salt to the transport water to enhance the stress resistance of the fish. Importers may adopt proper acclimation procedure and allow fish to recover in low salinity water to reduce post‐shipment mortality. As the main bulk of post‐shipment mortality is stress‐mediated and occurs during the 1‐week recovery period, the industry should consider revising the basis of the current warranty system for their customers, from death on arrival to cumulative mortality at 7 days post shipment (or death after 7 days, DA7), in order to cut down fish losses after shipment.     

Possible applications of modern fish larviculture technology to ornamental fish production

There has been rapid development in the marine foodfishlarviculture technology in Europe since the early eighties,especially in the flat fish, turbot and halibut, and the bass andbream species. The most significant improvements in the eightieswere the introduction of light control, artificial reproductiontechniques, appropriate water treatment and the use of rotifersand Artemia nauplii of specific sizes and in the late eightiesand early nineties the quality enhancement of live food organismsusing specific enrichment techniques. Present research is focusedon the implementation of several microbial techniques to improvethe hygiene of live prey and fish.Many of the modern larviculture techniques being used in marine foodfish could be adapted for application in the ornamental fish industry. For examples, research in the Onamental Fish Section, Primary Production Department, Singapore has demonstrated that the use of freshwater rotifers and Artemia nauplii would enable artificial rearing of Discus in the absence of the parent fish and improve the larval performance of Gouramis and Tetra larvae. The use of such small live food organisms is likely to facilitate breeding of new fish species with small larvae. Research has also indicated that the use of diets containing vitamin C and certain immuno-stimulants improved the stress resistance of guppy. Such techniques would have important application in the fish transport, an important aspect in the ornamental fish industry     

Feeding behavior under dark conditions in larvae of sutchi catfish <Emphasis Type="Italic">Pangasianodon hypophthalmus</Emphasis>

Sutchi catfish Pangasianodon hypophthalmus hatch with morphologically immature sensory organs; however, sensory organs develop rapidly with larval growth. Two-day-old larvae commenced ingesting Artemia nauplii. The larvae displayed many taste buds on the barbels, the head surface, and in the buccal cavity. Other sense organs were also well developed at this stage. Feeding experiments revealed that 2-day-old larvae ingested Artemia under both light and dark conditions, moreover, the larvae could ingest frozen dead Artemia. The ingestion rates for 4- and 7-day-old larvae were significantly higher under dark conditions than under light conditions. The rates using frozen dead Artemia were mostly higher than the rates using live Artemia. Therefore, feeding behavior under dark conditions is most likely not mediated by visual or mechanical senses, but rather by chemosensory senses, such as taste buds. Larval fish are vulnerable to predators; thus, if they can search for and eat food at night, they can avoid diurnal predators. The behavior observed here appears to represent their survival strategy. Moreover, these results suggest a new possibility that sutchi catfish larvae can be reared under dark or dim light conditions in order to improve survival and growth rates as in the case of African catfish Clarias gariepinus.     

Use of Diet Crossover to Determine the Effects of β‐glucan Supplementation on Immunity and Growth of Nile Tilapia,Oreochromis niloticus

Juvenile Nile tilapia were fed either a basal (control) diet (n = 6 aquaria) or a diet supplemented with 1 g/kg β‐glucan (n = 24 aquaria) for 4 wk. At the end of this period, fish receiving β‐glucan were continued on the same diet (n = 12 aquaria) or switched to the control diet (n = 12 aquaria) for 2 wk. After 6 wk, tilapia continuously fed the β‐glucan supplemented diets had improved weight gain and feed efficiency than those fed the control diet uninterrupted or switched from the β‐glucan diet to the control after 4 wk. Feeding tilapia β‐glucan for 4 wk and then switching to the basal diet for 2 wk caused a significant increase in the respiratory burst of polymorphonuclear lymphocytes (17.77 × 10³ units/1000 white blood cells [WBC]) compared to catfish fed the control diet (13.50 × 10³ units/1000 WBC) or the β‐glucan diet continuously (13.57 × 10³ units/1000 WBC), but other immune parameters were unaffected. Tilapia were then challenged with Streptococcus iniae. The two groups were divided again (n = 6 aquaria) postchallenge and continued on the same diet or switched to the other diet (β‐glucan or control) for another 3 wk. No differences in survival to S. iniae infection occurred between dietary groups.     

ECOLOGY OF MYCOPLASMAS IN CLINICALLY HEALTHY CATS

Mycoplasmas were isolated from various sites and organs of a series of 319 clinically healthy cats. They include M. felis, M. gateae, M. Arginini, A. laidlawii, feline ureaplasmas, M. pulmonis, M. arthritidis, and M. gallisepticum. In addition, there were 10 strains of mycoplasmas which could not be identified with the specific antiserums by growth inhibition tests. Antibody against M. felis was demonstrated by haemagglutination-inhibition and complement-fixation tests in cats which were over 6 months of age. However, no such antibody against M. felis was detected in animals which were less than 6 months old. No antibody against A. laidlawii, M. gateae, M. arginini and feline ureaplasmas was demonstrated by the same serological methods. The significance of these mycoplasmas in cats is described.     

Improved monospermic fertilization and subsequent blastocyst formation of bovine oocytes fertilized in vitro in a medium containing NaCl of decreased concentration

The objective of this study was to evaluate whether changes in NaCl concentration in a fertilization medium could improve normal fertilization and preimplantation development of bovine oocytes. In vitro matured bovine oocytes were inseminated with frozen-thawed semen for 18 hr in a Tyrode's medium with albumin, lactate and pyruvate (TALP), to which 114 (TALP-114), 96 (TALP-96) or 78 (TALP-78) mM NaCl was added. Presumptive zygotes were cultured for 192 hr in a modified TALP containing 90 mM NaCl, 1.5 mM glucose, 0.3% (w/v) BSA, minimal essential medium (MEM) essential and nonessential amino acids, and insulin-transferrin-selenium complex. Lower polyspermy rate was obtained by the insemination in TALP-96 (7.8 +/- 2.3%) than by the insemination in TALP-114 (25.6 +/- 1.4%), without decrease in male pronucleus (MPN) formation. Fertilization in TALP-78 also yielded decreased polyspermic fertilization (3.8 +/- 1.5%), but significant decrease in MPN formation was found (63.1 +/- 3.1%). In preimplantation development, more blastocysts developed from oocytes inseminated in TALP-96 (24.1 +/- 1.7%) than from oocytes inseminated in TALP-114 (16.8 +/- 1.4%). TALP-78, however, did not improve preimplantation development beyond the 8-cell stage compared with TALP-114. Mean cell number of blastocyst was higher when oocytes were fertilized in TALP-96 (137.0 +/- 4.5) than in TALP-114 (123.1 +/- 5.1) and in TALP-78 (102.3 +/- 4.5). These results demonstrate that insemination of bovine oocytes in a TALP with decreased NaCl concentration (96 mM) improves blastocyst formation and embryo viability. Decrease in NaCl concentration below 96 mM, however, may delay or inhibit MPN formation, and inhibits subsequent development in vitro.     

Risks to the aquatic ecosystem from the application of Metarhizium anisopliae for locust control in Australia

Laboratory tests of Metarhizium anisopliae var acridum Driver & Milner, at a dose of 1.3 x 10(6) conidia ml-1, had no adverse effects on nymphs of mayfly, Ulmerophlebia sp or 8-week-old fry of the rainbow fish, Melanotaenia duboulayi Castelnau. This dose was toxic to the cladoceran, Ceriodaphnia dubia Richard, causing 100% mortality in 48 h. When this test was repeated at doses of up to 6.7 x 10(3) conidia ml-1, there was only 5% mortality after 192 h. Spraying of artificial water sources with a very high dose of the fungus as an aqueous spray resulted in 80-130 conidia ml-1 at 15 cm depth in the first 24 h after spraying. The conidia rapidly settled out and were absent from the top 15 cm layer of water after about 50 h. A similar experiment using the oil formulation as used in field control resulted in a 2- to 20-fold lower level of conidia in the water. Finally, sampling actual water sources in spray areas revealed a very low level of contamination of the water, with a maximum mean level of 29 conidia ml-1 in the first 24 h after treatment. Thus the level of conidia likely to enter water during control campaigns is a small fraction of that required to kill cladocerans, the only sensitive non-target organism tested. It is concluded that the biopesticide is very unlikely to pose any hazard to aquatic organisms.     

Toxoplasma gondii in an African crested porcupine (Hystrix cristata).

An adult female crested porcupine (Hystrix cristata) was evaluated for acute onset of neurologic signs including head tilt, circling, and ataxia. She was found dead in her holding area 2 days after initially exhibiting clinical signs. Necropsy was unremarkable. Histopathology of brain tissue revealed the presence of protozoal cysts associated with inflammation as the underlying cause of clinical signs and death. Immunohistochemical staining of brain tissue for Toxoplasma gondii was strongly positive. PCR on fresh brain confirmed T. gondii as the causative organism. An adult male in the same enclosure has demonstrated similar neurologic signs over the past 3 years and has failed to respond to various medical treatments. Clinical disease associated with T. gondii has not been previously reported in this porcupine species or any other Old World porcupines, although there are several reports of clinical toxoplasmosis involving New World porcupine species.     

Soybean mosaic virus Helper Component-Protease Alters Leaf Morphology and Reduces Seed Production in Transgenic Soybean Plants

ABSTRACT Transgenic soybean (Glycine max) plants expressing Soybean mosaic virus (SMV) helper component-protease (HC-Pro) showed altered vegetative and reproductive phenotypes and responses to SMV infection. When inoculated with SMV, transgenic plants expressing the lowest level of HC-Pro mRNA and those transformed with the vector alone initially showed mild SMV symptoms. Plants that accumulated the highest level of SMV HC-Pro mRNA showed very severe SMV symptoms initially, but after 2 weeks symptoms disappeared, and SMV titers were greatly reduced. Analysis of SMV RNA abundance over time with region-specific probes showed that the HC-Pro region of the SMV genome was degraded before the coat protein region. Transgenic soybean plants that expressed SMV HC-Pro showed dose-dependent alterations in unifoliate leaf morphologies and seed production where plants expressing the highest levels of HC-Pro had the most deformed leaves and the lowest seed production. Accumulation of microRNAs (miRNAs) and mRNAs putatively targeted by miRNAs was analyzed in leaves and flowers of healthy, HC-Pro-transgenic, and SMV-infected plants. Neither expression of SMV HC-Pro nor SMV infection produced greater than twofold changes in accumulation of six miRNAs. In contrast, SMV infection was associated with twofold or greater increases in the accumulation of four of seven miRNA-targeted mRNAs tested.     

The effects of enrofloxacin on canine tendon cells and chondrocytes proliferation <Emphasis Type="Italic">in vitro</Emphasis>

Enrofloxacin, a fluoroquinolone antibiotic has been used widely in humans and domestic animals, including dogs, because of its broad-spectrum activity and relative safety. The side effects of fluoroquinolone, induced tendinopathy, tendonitis, spontaneous tendon rupture and cartilage damage, remain incompletely understood. In the present study, we investigated the in vitro effects of enrofloxacin on cell proliferation and induction of apoptosis in canine Achilles tendon cells and chondrocytes. Cell growth and proliferation after treating with enrofloxacin for 2–6 days was quantified by a colorimetric 2,3-bis{2-methoxy-4-nitro-5-sulfophenyl}-2H-tetrazolium-5-carboxyanilide inner salt (XTT) assay. The results showed that enrofloxacin could inhibit the proliferation of canine tendon cells and chondrocytes at increasing concentrations (10–200 μg/ml). The inhibition of proliferation of canine tendon cells and chondrocytes after exposure to enrofloxacin were associated with induction of apoptosis, as evidenced by the typical nuclear apoptotic condensed nuclei found using Hoechst 33258 staining. It was demonstrated that canine tendon cells and chondrocytes treated with 200 μg/ml enrofloxacin for 4 days exhibited apoptotic features and fragmentation of DNA. Enrofloxacin also increased the apoptosis of canine tendon cells and chondrocytes in a dose and time-dependent manner. The results indicate that enrofloxacin inhibits cell proliferation, induces apoptosis and DNA fragmentation, which might explain enrofloxacin-induced tendinopathy and cartilage damage.