Intercohort cannibalism and parturition-associated behaviour of adult swordtail, Xiphophorus helleri Heckel (Pisces: Poeciliidae)

The post‐parturition behavioural patterns of adult Xiphophorus helleri (Heckel) were described and quantified by recording their duration and/or frequency in glass tanks. All births occurred in the dark. Female behaviour might increase the chance of newborn juvenile survival: at the top of the tank, where juveniles are more vulnerable to predation, the females gave birth in the artificial refuge, while no parturition took place in the refuge at the bottom. Birth‐giving females were no less cannibalistic than males and other females. There was a reduction in cannibalism and the frequency of secondary attacks with time, even though the number of juveniles in the tank continued to increase due to ongoing parturition. Most adults were found at the bottom of the water column during the first 500 min of parturition, after which they were distributed equally in the water column. The movement of adults, away from the bottom and consequently away from the juveniles corresponded with an increase in feeding on alternative food and a reduction in cannibalism. Thus, cannibalism in swordtail might be opportunistic predation.     

Large-scale turbidostat culture of marine microalgae

A simple and inexpensive device is described which maintains cultures of marine microalgae at predetermined cell densities by the automatic control of harvesting and replenishment of the culture volume with fresh medium. Cultures were maintained at a range of cell densities in 80-litre internally illuminated vessels and the daily cell production yield was evaluated.Maximum yields of 0·8–1·0×10¹² cells day^?1 of Isochrysis aff. galbana Green and 0·8–1·2×10¹¹ cells day^?1 of Tetraselmis suecica (Kylin) Butch. were obtained at culture densities of 15–17×10³ cells μlitre^?1 and 19–21×10² cells μlitre^?1, respectively. Turbidostat cultures of Skeletonema costatum (Grev.) Cleve could not consistently be maintained above 5–7×10³ cells μlitre^?1.     

A Large‐scale,Rapid Public Health Response to Rabies in an Organ Recipient and the Previously Undiagnosed Organ Donor

This article describes and contrasts the public health response to two human rabies cases: one organ recipient diagnosed within days of symptom onset and the transplant donor who was diagnosed 18 months post‐symptom onset. In response to an organ‐transplant‐related rabies case diagnosed in 2013, organ donor and recipient investigations were conducted by multiple public health agencies. Persons with potential exposure to infectious patient materials were assessed for rabies virus exposure. An exposure investigation was conducted to determine the source of the organ donor's infection. Over 100 persons from more than 20 agencies spent over 2700 h conducting contact investigations in healthcare, military and community settings. The 564 persons assessed include 417 healthcare workers [5.8% recommended for post‐exposure prophylaxis (PEP)], 96 community contacts (15.6% recommended for PEP), 30 autopsy personnel (50% recommended for PEP), and 21 other persons (4.8% recommended for PEP). Donor contacts represented 188 assessed with 20.2% recommended for PEP, compared with 5.6% of 306 recipient contacts recommended for PEP. Human rabies cases result in substantial use of public health and medical resources, especially when diagnosis is delayed. Although rare, clinicians should consider rabies in cases of encephalitis of unexplained aetiology, particularly for cases that may result in organ donation.     

Duration and method of fixation affects the sensitivity of a digoxygenin-labelled DNA probe in detecting Kudoa thyrsites in Atlantic salmon skeletal muscle

The effect of fixative and duration of fixation on the sensitivity of a non-radioactive in situ hybridisation (ISH) protocol to detect Kudoa thyrsites small subunit ribosomal deoxyribonucleic acid (DNA) was investigated. Strong ISH reactions were detected in 5-μm sections of paraffin-embedded Atlantic salmon muscle after fixation for 1 day in Davidson's solution (DS). Reactions were weak following 3 or 5 days fixation and absent after 17 or 28 days fixation. Strong ISH reactions were observed after 1, 3 or 5 days fixation in neutral buffered formalin (NBF). The reactions were weak after 17 days and weak to nonexistent after 28 days of fixation. Reactions were consistently strong after fixation in 95% ethanol for up to 28 days. Some mature spores reacted weakly or not at all by ISH. Parasite DNA was weakly amplified by polymerase chain reaction (PCR) from paraffin-embedded muscle after 1 day of fixation in DS but not after fixation for 3, 5, 17 or 28 days. Amplified DNA was detected after fixation in NBF for 1, 3 and 5 days, but not after 17 or 28 days. In contrast, PCR consistently amplified DNA from paraffin-embedded, ethanol-fixed muscle. Caution should be used in the choice of fixative and duration of fixation when preserving Atlantic salmon tissues for molecular diagnosis of K. thyrsites.     

The effect of dietary protein source on growth and carcass composition in juvenile Australian freshwater crayfish

A feed trial was conducted for 12 weeks on juvenile Australian freshwater crayfish (Cherax destructor) (mean weight (SE) 0.82 (0.02)g) maintained on five isoenergetic diets with a protein content of 30%. Diets differed in the primary source of protein used, with meat, snail, soybean, yabby, and zooplankton meals comprising the major protein ingredient, varying from 56–60% of total protein. Mean percentage weight gain per day ranged from 7.57% (yabby meal diet), to 9.42% (snail meal diet). No significant difference occurred in mean weight, percentage weight gain, specific growth rate (%), or survival among diets. A maximum size of 16.44g was attained on the yabby meal diet. Largest mean weight was 8.27g on the snail-based diet. Food conversion ratios were all good, with a minimum value of 0.95 recorded for the snail-based diet. Initial food consumption per day was approximately 5% of mean animal weight, decreasing to around 2.4%, and is collectively described by a power curve. Protein retention ranged from 29.57% in the zooplankton meal diet to 41.15% in the snail-based diet. Carcass composition was influenced by feed type, with the most marked difference occurring in carapace colour. Animals fed the zooplankton-based pellets developed the strongest pigmentation. Recommendations are made for including certain protein-based ingredients in manufactured yabby diets.     

Characterization of serum and mucosal antibody responses and relative per cent survival in rainbow trout, Oncorhynchus mykiss (Walbaum), following immunization and challenge with Flavobacterium psychrophilum

Serum and mucosal antibody responses of juvenile rainbow trout, Oncorhynchus mykiss, were characterized by enzyme‐linked immunosorbent assay (ELISA) following immunization with various preparations of formalin‐killed Flavobacterium psychrophilum cells. The protective nature of these preparations was then determined by immunizing rainbow trout fry and challenging with the bacterium. Juvenile rainbow trout immunized intraperitoneally (i.p.) with formalin‐killed F. psychrophilum emulsified with Freund's complete adjuvant (FCA), and i.p. with formalin‐killed F. psychrophilum either with or without culture supernatant generated significant serum antibody responses by 6 and 9 weeks, respectively. Significant mucosal antibody responses were detected by 9 weeks only in fish immunized i.p. with killed F. psychrophilum/FCA. Following immunization and bacterial challenge of rainbow trout fry, protective immunity was conferred in F. psychrophilum/FCA and saline/FCA groups with relative per cent survival values of up to 83 and 51, respectively. Significant protection was not observed in treatment groups immunized by immersion or i.p. without adjuvant at the challenge doses tested. Results suggest that stimulation of non‐specific immune factors enhances the ability of fish to mount a protective immune response, but specific antibody appears necessary to provide near complete protection. In this study, an ELISA was developed to monitor anti‐F. psychrophilum antibody production in trout. The relationship of such responses to protective immunity suggests that future vaccination strategies against coldwater disease may require stimulation of both the innate and adaptive arms of the immune response.     

Trypsin enzyme activity during larval development of Litopenaeus vannamei (Boone) fed on live feeds

Larval stages of the Pacific white shrimp, Litopenaeus vannamei (Boone) were fed standard live diets of mixed microalgae from the first to the third protozoea (PZ1 to PZ3), followed by Artemia nauplii until post‐larvae 1 (PL1). Trypsin enzyme activity for each larval stage was determined using N‐α‐p‐toluenesulphonyl‐l ‐arginine methyl ester (TAME) as a substrate. Results were expressed as enzyme content to assess ontogenetic changes during larval development. Tissue trypsin content (IU µg^?1 DW for each larval stage) was significantly highest at the PZ1 stage and declined through subsequent stages to PL1. This contrasts with previously observed patterns of trypsin development in Litopenaeus setiferus (Linnaeus) and other penaeid genera, which exhibit a peak in trypsin activity at the third protozoea/first mysis (PZ3/M1) larval stage. Litopenaeus vannamei larvae transferred to a diet of Artemia at the beginning of the second protozoea (PZ2) stage were significantly heavier on reaching the first mysis stage (M1) than those fed algae, while survival was not significantly different between treatments. At both PZ2 and PZ3 stages, trypsin content in larvae feeding on Artemia was significantly lower than in those feeding on algae. The rapid decline in trypsin content from PZ1 and the flexible enzyme response from PZ2 suggest that L. vannamei is physiologically adapted to transfer to a more carnivorous diet during the mid‐protozoeal stages.     

The efficiency and condition of oysters and macroalgae used as biological filters of shrimp pond effluent

Current shrimp pond management practices generally result in elevated concentrations of nutrients, suspended solids, bacteria and phytoplankton compared with the influent water. Concerns about adverse environmental impacts caused by discharging pond effluent directly into adjacent waterways have prompted the search for cost‐effective methods of effluent treatment. One potential method of effluent treatment is the use of ponds or raceways stocked with plants or animals that act as natural biofilters by removing waste nutrients. In addition to improving effluent water quality prior to discharge, the use of natural biofilters provides a method for capturing otherwise wasted nutrients. This study examined the potential of the native oyster, Saccostrea commercialis (Iredale and Roughley) and macroalgae, Gracilaria edulis (Gmelin) Silva to improve effluent water quality from a commercial Penaeus japonicus (Bate) shrimp farm. A system of raceways was constructed to permit recirculation of the effluent through the oysters to maximize the filtration of bacteria, phytoplankton and total suspended solids. A series of experiments was conducted to test the ability of oysters and macroalgae to improve effluent water quality in a flow‐through system compared with a recirculating system. In the flow‐through system, oysters reduced the concentration of bacteria to 35% of the initial concentration, chlorophyll a to 39%, total particulates (2.28–35.2 µm) to 29%, total nitrogen to 66% and total phosphorus to 56%. Under the recirculating flow regime, the ability of the oysters to improve water quality was significantly enhanced. After four circuits, total bacterial numbers were reduced to 12%, chlorophyll a to 4%, and total suspended solids to 16%. Efforts to increase biofiltration by adding additional layers of oyster trays and macroalgae‐filled mesh bags resulted in fouling of the lower layers causing the death of oysters and senescence of macroalgae. Supplementary laboratory experiments were designed to examine the effects of high effluent concentrations of suspended particulates on the growth and condition of oysters and macroalgae. The results demonstrated that high concentrations of particulates inhibited growth and reduced the condition of oysters and macroalgae. Allowing the effluent to settle before biofiltration improved growth and reduced signs of stress in the oysters and macroalgae. A settling time of 6 h reduced particulates to a level that prevented fouling of the oysters and macroalgae.