Quantitative Growth of Naegleria in Axenic Culture

A strain of Naegleria gruberi, isolated from a Vero cell culture and designated TS-1, was axenically cultivated in monolayer and mass aerating suspension culture. Cultural conditions for constant growth parameters and high-exponential cell densities were defined. Serum or other supplemented fractions were found essential in both Trypticase-yeast extract-glucose (TYG) and Casitone (CAS)-based media. Monolayer cultures grown in the CAS medium required lower levels of serum to reach maximum stationary densities of amoebae than cultures grown in the TYG medium. Heat-killed (121 C, 10 min) whole cell and cell lysate bacterial fractions were capable of replacing the serum in both the TYG and CAS media. Heat-killed bacterial fractions provided the same levels of growth as attained with serum in TYG medium, whereas the bacterial lysate supported only minimal growth in the same medium. In the CAS medium, both bacterial fractions resulted in the same level of growth which was equal to that obtained in reduced serum content. Strain TS-1 was established in suspension culture with the CAS medium used in monolayer culture. The addition of sheep red blood cells (RBC) or RBC lysate greatly enhanced growth responses. Further modifications resulted in a final medium for suspension culture consisting of Casitone-yeast extract-glucose-vitamin base, supplemented with serum and RBC lysate. This medium supported growth with a mean generation time of 9 h at 30 C and a stationary phase yield of greater than 5 x 10(6) amoebae per ml.     

Courtship and mating behaviour of manta rays Mobula alfredi and M. birostris in the Maldives

The aim of this 14 year study was to elucidate the entire courtship and mating behaviour of manta rays Mobula alfredi and M. birostris using behavioural observations, video and photographic records. From 2003 to 2016, over 11,000 surveys were undertaken at known manta ray aggregation sites in the Maldives to record any observed manta rays reproductive activity. From 47,591 photo‐ID sightings, 4,247 individual M. alfredi were identified and 226 individual M. birostris from 229 photo‐ID sightings, all recorded at 22 atolls across 265 different sites. Courtship activity was observed on 206 surveys at 30 different sites. A total of 229 courtship events were recorded, with 90% (n = 205) of them occurring at cleaning sites. The observed courtship activity was categorised into seven distinct stages which are described in detail: initiation, endurance, evasion, pre‐copulatory positioning, copulation, post‐copulatory holding and separation. Photographs provide the first scientific record of the entirety of manta rays courtship and mating. Both M. alfredi and M. birostris appear to engage in the same elaborate courtship rituals, exhibiting the same behaviours during all stages of the courtship and mating process.     

Evaluation of tissue changes following intramuscular infiltration of lidocaine in rainbow trout Oncorhynchus mykiss

Rainbow trout Oncorhynchus mykiss were infiltrated with either saline or lidocaine adjacent to the dorsal fin to assess histopathological changes. Infiltration was done as if it were being used as a local anaesthetic. Tissue lesions and associated tissue healing were examined over a period of 30 days. Most changes occurred at the cranial site of where the solution was first infiltrated. The infiltration of a dose of 10 mg kg^?1 of lidocaine appears to have damaged the skeletal muscle and connective tissues more than a similar volume of saline, especially during the first 15 days. The primary changes included haemorrhage, inflammation and muscle degeneration and necrosis. By day 30 post‐infiltration inflammatory lesions were either nearly or completely absent, signs of myofibre regeneration were noted in only one fish. This experiment shows local anaesthetics and saline can produce localized tissue damage, especially during the first 2 weeks post infiltration. Care should be taken to allow the fish to heal for at least 30 days and probably more, no matter the solution administered, especially if giving repeated injections or infiltrations at the same site.     

Hyperstability masks declines in bumphead parrotfish (<Emphasis Type="Italic">Bolbometopon muricatum</Emphasis>) populations

Bolbometopon muricatum, the largest species of parrotfish, is a functionally important species that is characterised by the formation of aggregations for foraging, reproductive, and sleeping behaviours. Aggregations are restricted to shallow reef habitats, the locations of which are often known to local fishers. Bolbometopon muricatum fisheries are therefore vulnerable to overfishing and are likely to exhibit hyperstability, the maintenance of high catch per unit effort (CPUE) while population abundance declines. In this study, we provide a clear demonstration of hyperstable dynamics in a commercial B. muricatum fishery in Isabel Province, Solomon Islands. Initially, we used participatory mapping to demarcate the Kia fishing grounds into nine zones that had experienced different historic levels of fishing pressure. We then conducted comprehensive underwater visual census (UVC) and CPUE surveys across these zones over a 21-month period in 2012–2013. The individual sites for replicate UVC surveys were selected using a generalised random tessellation stratified variable probability design, while CPUE surveys involved trained provincial fisheries officers and local spearfishers. A comparison of fishery-independent abundance data and fishery-dependent CPUE data indicate extreme hyperstability, with CPUE maintained as B. muricatum abundance declines towards zero. Hyperstability may explain the sudden collapses of many B. muricatum spear fisheries across the Pacific and highlights the limitations of using data-poor fisheries assessment methods to evaluate the status of commercially valuable coral reef fishes that form predicable aggregations.     

Gibbon Aggression During Introductions: An International Survey

Little is known regarding the prevalence of aggression seen during introductions of captive gibbons (Hylobatidae). In this study, an online survey was developed to quantify and collect contextual details regarding the frequency and types of aggression seen during introductions of captive gibbons (Hylobatidae). Nineteen percent of institutions (17 institutions) reported observing aggression, and 6 of these institutions recorded multiple instances of aggression, though a vast majority of these cases resulted in mild injuries or none at all. The female was the primary aggressor in 23% of cases, the male was the primary aggressor in 58% of cases, and both were the primary aggressor in 1 case. Although these aggressive interactions were often not associated with a known cause, 27% of cases were associated with food displacement. In most cases, management changes, including trying new pairings, greatly reduced situational aggression, suggesting that individual personalities may play a factor in aggression. These data begin to explain the extent of aggression observed in captive gibbons; future studies will address possible correlations with aggression and introduction techniques.