Classification of guppies’ (Poecilia reticulata) gender by computer vision

Labor costs of guppy growers and breeders are largely those of manual sorting (by strain, quality and gender) and counting fish. In most farms, female and male fish are grown together and sold either separately or together. Sorting fish according to gender is important for marketing as well as for breeding programs, so that a device for sorting and counting fish can potentially reduce production costs and improve quality.

A project aiming to develop sorting and counting technologies for ornamental fish growers included development and testing of image-processing algorithms for sorting guppy fish (Poecilia reticulata) by gender. The algorithms are derived from shape and color differences between female and male guppies. An algorithm for the determination of landmarks on fish contours was developed and found to be accurate in accordance with human judgment, enabling extraction of specific shape and color features of the tail and the body.

The algorithms were applied to three sets of images of guppies of the “Red-Blond” strain. Gender identification accuracy was approximately 90% using shape features, approximately 96% using color features and was slightly improved when both color and shape features were used.

Some of the components used are essential for future development of a computer vision based system for sorting and grading ornamental fish by strain and quality.     

Solution of a protein crystal structure with a model obtained from NMR interproton distance restraints

Model calculations were performed to test the possibility of solving crystal structures of proteins by Patterson search techniques with three-dimensional structures obtained from nuclear magnetic resonance (NMR) interproton distance restraints. Structures for crambin obtained from simulated NMR data were used as the test system; the root-mean-square deviations of the NMR structures from the x-ray structure were 1.5 to 2.2 A for backbone atoms and 2.0 to 2.8 A for side-chain atoms. Patterson searches were made to determine the orientation and position of the NMR structures in the unit cell. The correct solution was obtained by comparing the rotation function results of several of the NMR structures and the average structure derived from them. Conventional refinement techniques reduced the R factor from 0.43 at 4 A resolution to 0.27 at 2 A resolution without inclusion of water molecules. The partially refined structure has root-mean-square backbone and side-chain atom deviations from the x-ray structure of 0.5 and 1.3 A, respectively.     

Shifting the natural spring–summer breeding season of the Australian freshwater crayfish Cherax quadricarinatus into the winter by environmental manipulations

Ninety sexually mature Cherax quadricarinatus females were exposed to various combinations of photoperiod and temperature for 2 months during the summer. Females were randomly assigned to either “winter” “semi-winter” or “summer” simulation treatments. In the “winter” treatment, crayfish were exposed to a simulated winter photoperiod (gradual decrease from 14L:10D to 10L:14D, 4 weeks at short day length followed by gradual increase to 14L:10D) and temperature (gradual decrease from 27 to 15 °C, held for 4 weeks, and then gradual increase to 27 °C). In the “semi-winter” treatment, crayfish were exposed to a simulated winter photoperiod and a summer temperature (27–29 °C). In the “summer” treatment, the crayfish were exposed to summer water temperatures and a photoperiod of 14L:10D. Following the 2 months of conditioning, the females were stocked for 7 months in small groups with males under environmental conditions similar to those of the “summer” treatment. All females were individually tagged and molting and spawning events were monitored. Females exposed to “semi-winter” conditioning in the summer, demonstrated a threefold increase in the rate of first spawning during the winter (December–February) compared with the other females. Crayfish breeders can easily implement these findings since shifting the breeding season into the winter only requires shortening of the photoperiod in the summer. The stocking of ponds in the spring with large nursed juveniles that hatched from eggs spawned in the winter, would allow the attainment of market size at the end of the limited growout season in temperate zones.     

Guidance of groups of guppies (Poecilia reticulata) to allow sorting by computer vision

Prior to marketing, ornamental tropical fish are currently sorted, graded and counted manually; a process that is labor intensive, stressful to fish and inaccurate. Manipulation of fish movements at our will is a key element in the development of a device for automatically sorting, grading and counting fish by a computer vision system, and the present study is one step towards the development of such a fish-friendly sorting device. The positive phototactic and rheotactic innate responses of guppies (Poecilia reticulata) were exploited to induce them to swim through transparent pipes or narrow channels, to render them ready for inspection by a computer vision system. Water flow direction and velocity, water level, and illumination were effective in inducing fish to move from one container into the other via a narrow transparent pipe. It was attempted to separate guppies moving in groups into individually spaced fish, by introducing an obstacle into a narrow channel or narrowing the transparent pipe.     

Epitheliocystis infection in wild and cultured sea bream (Sparus aurata, Sparidae) and grey mullets (Liza ramada, Mugilidae)

Heavy infection with epitheliocystis on gills caused mortality in hatchery-born Sparus aurata. Histopathological findings showed extensive proliferation of the gill epithelium around the epitheliocystis capsule. Such proliferation was not observed in benign infections. Epizootic infections thus occurred only in hatchery-born S. aurata. In S. aurata and mullets reared from natural stocks, infection remained benign and sporadic. Epitheliocystis infections were also found in juvenile mullets from the east Mediterranean waters as well as from the Gulf of Elat, Red Sea.     

Anatomy of a conformational change: hinged "lid" motion of the triosephosphate isomerase loop

Triosephosphate isomerase (TIM) is used as a model system for the study of how a localized conformational change in a protein structure is produced and related to enzyme reactivity. An 11-residue loop region moves more than 7 angstroms and closes over the active site when substrate binds. The loop acts like a "lid" in that it moves rigidly and is attached by two hinges to the remainder of the protein. The nature of the motion appears to be built into the loop by conserved residues; the hinge regions, in contrast, are not conserved. Results of molecular dynamics calculations confirm the structural analysis and suggest a possible ligand-induced mechanism for loop closure.     

Hidden thermodynamics of mutant proteins: a molecular dynamics analysis

A molecular dynamics simulation method is used to determine the contributions of individual amino acid residues and solvent molecules to free energy changes in proteins. Its application to the hemoglobin interface mutant Asp G1(99) beta----Ala shows that some of the contributions to the difference in the free energy of cooperativity are as large as 60 kilocalories (kcal) per mole. Since the overall free energy change is only -5.5 kcal/mole (versus the experimental value of -3.4 kcal/mole), essential elements of the thermodynamics are hidden in the measured results. By exposing the individual contributions, the free energy simulation provides new insights into the origin of thermodynamic changes in mutant proteins and demonstrates the role of effects beyond those usually considered in structural analyses.     

Parasitism of <Emphasis Type="Italic">Trichoderma</Emphasis> on <Emphasis Type="Italic">Meloidogyne javanica</Emphasis> and role of the gelatinous matrix

Trichoderma (T. asperellum-203, 44 and GH11; T. atroviride-IMI 206040 and T. harzianum-248) parasitism on Meloidogyne javanica life stages was examined in vitro. Conidium attachment and parasitism differed beween the fungi. Egg masses, their derived eggs and second-stage juveniles (J2) were parasitized by Trichoderma asperellum-203, 44, and T. atroviride following conidium attachment. Trichoderma asperellum-GH11 attached to the nematodes but exhibited reduced penetration, whereas growth of T. harzianum-248 attached to egg masses was inhibited. Only a few conidia of the different fungi were attached to eggs and J2s without gelatinous matrix; the eggs were penetrated and parasitized by few hyphae, while J2s were rarely parasitized by the fungi. The gelatinous matrix specifically induced J2 immobilization by T. asperellum-203, 44 and T. atroviride metabolites that immobilized the J2s. A constitutive-GFP-expressing T. asperellum-203 construct was used to visualize fungal penetration of the nematodes. Scanning electron microscopy revealed the formation of coiling and appressorium-like structures upon attachment and parasitism by T. asperellum-203 and T. atroviride. Gelatinous matrix agglutinated T. asperellum-203 and T. atroviride conidia, a process that was Ca²⁺-dependent. Conidium agglutination was inhibited by carbohydrates, including fucose, as was conidium attachment to the nematodes. All but T. harzianum could grow on the gelatinous matrix, which enhanced conidium germination. A biomimetic system based on gelatinous-matrix-coated nylon fibers demonstrated the role of the matrix in parasitism: T. asperellum-203 and T. atroviride conidia attached specifically to the gelatinous-matrix-coated fibers and parasitic growth patterns, such as coiling, branching and appressoria-like structures, were induced in both fungi, similarly to those observed during nematode parasitism. All Trichoderma isolates exhibited nematode biocontrol activity in pot experiments with tomato plants. Parasitic interactions were demonstrated in planta: females and egg masses dissected from tomato roots grown in T. asperellum-203-treated soil were examined and found to be parasitized by the fungus. This study demonstrates biocontrol activities of Trichoderma isolates and their parasitic capabilities on M. javanica, elucidating the importance of the gelatinous matrix in the fungal parasitism.     

Isospora carliae sp. n. (Apicomplexa: Eimeriidae) from the skink Carlia rhomboidalis (Peters) from Daintree Forest, North Queensland, Australia: description and fine-structural account of endogenous development

Isospora carliae sp. n. is described from the blue-throated rainbow skink Carlia rhomboidalis (Peters), from Daintree Forest, North Queensland, Australia. Oocysts are ellipsoidal, 16.8-21.0 x 12.6-15.4 microm in size, with their two sporocysts, 9.0-14.0 x 7.0-9.24 microm in size, positioned along the wide axis. Sporozoites contain a distinct refractile body and are accompanied by a residuum. All endogenous development occurs within the host-cell nucleus. Nuclei are sometimes invaded by several merozoites, but only infections by a single parasite persist. Nuclei lodging meronts, mature microgamonts and premature macrogamonts have an elongate shape. Some meronts exhibit a membrane-bound cytoplasmic inclusion that contains many micronemes.