Refrigerated Storage of Channel Catfish Sperm

Abstract.— Refrigerated storage of sperm is useful for genetic study and artificial breeding of fishes. Due to the potential loss of donor males, storage is important in species such as channel catfish Icralurus punctatus from which sperm cannot be stripped. This study addresses short-term storage (at 4 C) of channel catfish sperm by evaluation of storage methods employed for other species and for cryopreservation of channel catfish sperm. The objectives were to evaluate: 1) storage of intact testes and storage of sperm suspended in an extender solution; 2) use of various storage containers with and without supplemental oxygen; 3) use of extender solution with and without the addition of an antibiotic/antimycotic cocktail; 4) use of extender solution with and without the addition of methanol; and 5) use of extender solution with and without the addition of glucose and methanol. Sperm suspended in extender solution retained motility significantly longer (9 d) than did sperm in intact testis (2 d). Sperm stored in Zip-loc® plastic bags inflated with pure oxygen retained motility significantly longer (12 d) than did sperm stored in Zip-loc® plastic bags without supplemental oxygen (7 d), or sperm stored in plastic beakers (8 d) or test tubes (8 d) without supplemental oxygen. Sperm stored with the addition of antibiotic/antimycotic cocktail or methanol retained motility significantly longer (10–12 d) than did sperm stored without additives (6–8 d). Sperm stored in extender solution without glucose retained motility significantly longer (19–21 d) than did sperm stored in extender with glucose (13–16 d). Motility was retained for as long as 21 d in sperm stored in extender solution with 5% methanol and without glucose. In each experiment, loss of motility was associated with bacterial growth.     

The exploitation of the genome in the search for determinants of virulence in Streptococcus uberis

Despite much success in the control of mastitis in dairy cattle, intramammary infection with Streptococcus uberis remains a threat to herd health. This organism is a frequent cause of mastitis worldwide. Recent advances in the ability to genetically manipulate this bacterium, coupled to the determination of a representative genome sequence have already enabled the investigation of certain aspects of disease pathogenesis. Further use of such technology coupled to reliable models of disease and post-genomic analysis will permit the elucidation of further interactions between pathogen and host. This additional information can be usefully targeted at identification of candidates for inclusion in effective vaccines. This communication reviews the current, reported progress using this technology for S. uberis.     

Structure and receptor specificity of the hemagglutinin from an H5N1 influenza virus

The hemagglutinin (HA) structure at 2.9 angstrom resolution, from a highly pathogenic Vietnamese H5N1 influenza virus, is more related to the 1918 and other human H1 HAs than to a 1997 duck H5 HA. Glycan microarray analysis of this Viet04 HA reveals an avian alpha2-3 sialic acid receptor binding preference. Introduction of mutations that can convert H1 serotype HAs to human alpha2-6 receptor specificity only enhanced or reduced affinity for avian-type receptors. However, mutations that can convert avian H2 and H3 HAs to human receptor specificity, when inserted onto the Viet04 H5 HA framework, permitted binding to a natural human alpha2-6 glycan, which suggests a path for this H5N1 virus to gain a foothold in the human population.     

Conservation of Pattern and Process: Developing an Alternative Paradigm of Rangeland Management

This article examines the question of how well the rangeland management profession has served conservation of patterns and processes that support multiple ecosystem services. We examine the paradigms under which rangeland management operates and argue that our profession developed under the utilitarian paradigm with the primary goals of sustainable forage for livestock production. While optimization of multiple rangeland products and services has always been a consideration, a comprehensive set of principles have not be been developed to advance this concept. We argue that fire and grazing, often viewed as mere tools used for production goals, should rather be viewed as essential ecosystem processes. Rangeland management continues to operate under the utilitarian paradigm appropriate to societal values of the 20th century and by and large has failed to provide management guidance to reverse degradation of several highly valued ecosystem services. We support this argument with evidence that biodiversity has declined on rangelands in the past half century and that much of this decline is due to management goals that favor a narrow suite of species. The full suite of ecosystem services valued by society will only benefit by management for heterogeneity, which implies that there is no one goal for management and that landscape-level planning is crucial. Explicitly incorporating heterogeneity into state-and-transition models is an important advancement not yet achieved by our profession. We present new principles for rangeland management formed on the basis of conservation of pattern and process. While recognizing that many rangelands have significant deviations from historic plant communities and disturbance regimes, we suggest that management for conservation of pattern and process should focus on fire and grazing to the extent possible to promote a shifting mosaic across large landscapes that include patches that are highly variable in the amount of disturbance rather than the current goal of uniform moderate disturbance.     

Evaluation of Commercial‐scale Approaches for Cryopreservation of White Crappie,Pomoxis annularis,Sperm

Crappie, Pomoxis spp., are popular game fish throughout North America and are produced by public and private hatcheries. However, production is limited by a lack of information on tank culture and induced spawning methods. Development of techniques for storage of sperm and in vitro fertilization would increase flexibility in spawning. Therefore, techniques for sperm cryopreservation were examined in white crappie, Pomoxis annularis. Sperm from adult wild white crappie were used to evaluate sperm extender, cryoprotectant agent and concentration, and cooling technique based on post‐thaw sperm motility. Percent egg fertilization was also compared between sperm stored in the two best cryopreservation protocols and two different osmotic activator solutions. Sperm were cryopreserved using treatment combinations of two extenders (350 mOsmol/kg Hanks' balanced salt solution [HBSS] and 350 mOsmol/kg Ca²⁺free HBSS) and two cryoprotectants (dimethyl sulfoxide [DMSO] and methanol) at concentrations of 5, 10, and 15% that were cooled at four different rates: 5, 10, 20, and 40 C/min. Post‐thaw sperm motility and fertilization rates indicated white crappie sperm can be cryopreserved using either extender, cryoprotectants of either 5% DMSO or 10% methanol, and cooling at 40 C/min. A follow‐up experiment demonstrated sperm in suspensions on ice retained viability after overnight transport.     

A real-time polymerase chain reaction assay for the detection of the myxozoan parasite Henneguya ictaluri in channel catfish

Proliferative gill disease (PGD), caused by the myxozoan parasite Henneguya ictaluri, is the most prevalent parasitic infection affecting commercial channel catfish (Ictalurus punctatus) aquaculture. There are currently no effective chemotherapeutic or biological control measures for PGD, which often peaks during the spring and fall when water temperatures are between 16-25 degrees C. The current diagnostic techniques of gross examination of gill clip wet mounts and histopathology are subject to false-negatives during the early stages of infection, and the quantifiable nature of end-point polymerase chain reaction (PCR) is subjective. Consequently, a rapid and more sensitive quantitative real-time PCR assay was developed for the detection of H. ictaluri during the early stages of infection in channel catfish. A 23 base-pair TaqMan probe was designed based on previously published H. ictaluri PCR protocols. The sensitivity of the assay was the equivalent of a single H. ictaluri actinospore, and in a pond challenge study, quantitative real-time PCR proved to be more sensitive than gross examination, microscopic examination of gill clip wet mounts, and histopathologic examination of gill tissue sections. Future applications of this assay will focus on developing methodologies to be used in conjunction with current pond-monitoring protocols to evaluate potential treatments and better manage this significant seasonal disease.     

Effects of reconcile (fluoxetine) chewable tablets plus behavior management for canine separation anxiety

Canine separation anxiety is a common behavioral problem presented to veterinarians. Associated behaviors are distressing to both dog and owner, have the potential to disrupt the human-companion animal bond, and may lead to euthanasia. The results of this study demonstrate the clinical efficacy and safety of Reconcile (fluoxetine, 1 to 2 mg/kg/day [0.45 to 0.91 mg/lb/day]), in conjunction with behavior management, for the treatment of canine separation anxiety. The beef flavored chewable formulation was palatable to treated dogs and easy to administer. This study provides to veterinarians and owners valuable information about an effective separation anxiety treatment plan that combines use of Reconcile with behavior modification.     

Environmental values and forest patch conservation in a rural Costa Rican community

Although conservation attention has generally focused on large forest tracts, there is increasing evidence that smaller forest patches are important for both conservation and rural development. A study of forest patch conservation in a rural Costa Rican community found that, although forest patch conservation was influenced by landholding size, material factors did not account for all the variation in forest patches conservation behavior or conservation orientations of farmers. A qualitative interpretive approach, using semi-structured interviews, found that environmental values were influenced by at least three factors. First, colonist farmers with direct experience with environmental degradation in other regions of Costa Rica were more likely to be concerned about environmental degradation. Second, childhood exposure to conservation played an important role in influencing conservation orientations. Third, the environment was frequently discussed in relation to the moral and social values present in religious doctrines. The study also found general support for forest conservation laws, and a clear indication that interest in forest conservation has grown in recent years. The study highlights the importance of environmental values, in conjunction with material factors, in influencing forest conservation in rural communities.     

Vitrification of sperm from marine fish: effect on motility and membrane integrity

Our goal was to develop a standardized approach for sperm vitrification of marine fish that can be applied generally in aquatic species. The objectives were to: (i) estimate acute toxicity of cryoprotectants over a range of concentrations; (ii) evaluate the properties of vitrification solutions (VS); (iii) evaluate different thawing solutions and (iv) evaluate sperm quality after thawing by examination of motility and membrane integrity. Sperm were collected from red snapper (Lutjanus campechanus), spotted seatrout (Cynoscion nebulosus) and red drum (Sciaenops ocellatus). A total of 29 combinations of cryoprotectants were evaluated for toxicity and glass formation. Samples were loaded onto 10‐μL polystyrene loops and plunged into liquid nitrogen. There was a significant difference (P < 0.05) in post‐thaw motility among VS and among species when using the same VS. The sperm in VS of 15% DMSO + 15% ethylene glycol + 10% glycerol + 1% X‐1000^? + 1% Z‐1000^? had an average post‐thaw motility of 58% and membrane integrity of 19% for spotted seatrout, 38% and 9% for red snapper, and 30% and 19% for red drum. Adaptations by marine fish to higher osmotic pressures could explain the survival in the high cryoprotectant concentrations. Vitrification offers an alternative to conventional cryopreservation.     

Design and development of a geothermal temperature control system for broodstock management of channel catfish Ictalurus punctatus

A control system was designed to raise and maintain water temperatures within 0.03-ha earthen ponds to a range conducive for spawning (24–30 °C) channel catfish Ictalurus punctatus. Heating was done during February 2001 to April 2001, when temperatures would have otherwise prohibited spawning (<24 °C). Temperature was increased from 10 °C (ambient) by 2 °C per day, and maintained at 27 °C, by the addition of geothermally warmed water (36 °C). The control system substantially increased the controllability and precision of heating ponds compared to manual operation. Systems were designed to control sets of four ponds. In designing this control system, consideration of biological constraints was essential. Reproduction in channel catfish is most strongly influenced by temperature. Because cold fronts are common during the winter and early spring (January–March), it was essential to ensure that pond temperatures did not fall below the range for spawning. Constraints on the heating rate and temperature variability to maintain fish health and stimulate spawning behavior were considered. Components of the control system included temperature measurement devices (type-T thermocouples), a central electronic control unit, electronic switches and electrically actuated ball valves. In response to the temperature sensed by each thermocouple, the controller sent a message to close or open the valve. When the valve was opened, warm water was added to the pond to increase the average pond temperature. Hardware and algorithm design and initial system testing were the major components of this project. The final design incorporated information on relevant biological parameters and safety features including peak pond temperature, independent aeration and water pressure control mechanisms. Initial results indicate successful control of this biological system, and ongoing studies suggest similar mechanisms may be used for additional control objectives. In particular, this system could be used to vary pond temperatures to study biological responses and to cool ponds by addition of well water during summer months.