Ecology and growth of juvenile California spiny lobster, Panulirus interruptus (Randall)

ABSTRACT TRANSCRIBED FROM ENGLE'S PH.D. ORAL DEFENSE PAMPHLET: The natural history of juvenile California spiny lobster, Panulirus interruptus (Randall), was investigated, with primary emphasis placed on ascertaining juvenile habitats, determining juvenile growth rates and component growth processes, and evaluating ecological and behavioral phenomena associated with juvenile survival and growth. Habitat surveys of island and mainland localities throughout southern and lower California revealed that small, greenish juveniles typically inhabit crevices or temporary burrows in 0-4m deep, wave-swept rocky habitats covered by dense beds of surf grass, Phyllospadix torreyi S. Watson. Phyllospadix beds were more abundant on gradually sloping rocky mainland beaches than on steeply sloping island shores. Phyllospadix abundance was positively correlated with P. interruptus abundance; however, at Santa Catalina Island, the Phyllospadix habitat was not extensive enough to be the sole lobster nursery. In laboratory tests, puerulus larvae and early juveniles chose Phyllospadix over rubble rocks or broad-bladed kelp, but did not consistently prefer Phyllospadix over reticulate algae. Ecology, growth, and behavior of juvenile P. interruptus inhabiting a discrete Phyllospadix habitat at Bird Rock, Santa Catalina Island, were investigated from October 1974 through December 1976 by means of frequent scuba surveys. Pueruli settled from June to November. Peak recruitment occurred from July to September, when seasonal temperatures were maximal. Settled larvae were approximately one year old. Juvenile growth was determined by size-frequency, single molt increment, mark-recapture, and laboratory culture studies. Carapace length vs. wet weight relationships fit standard power curve equations. Bird Rock juveniles grew from 7 to 32mm CL in 10-11 molts and from 32 to 56mm CL in 5-6 molts during their first and second benthic years, respectively. Growth rates were similar for males and females. Juveniles regenerating more than two limbs grew less per molt than intact lobsters. Long-term growth of laboratory-reared juveniles was 20% less than that of field lobsters. Growth component multiple regression analyses demonstrated that molt increment was directly proportional to premolt size and temperature for age 1+ lobsters. Molt frequency was inversely proportional to size and directly proportional to temperature. Temperature affected age 2+ lobsters similarly, but molt increment was independent of size, and molt frequency declined at a different rate. Juvenile growth rates more than doubled during warm water months compared to cold water months, primarily because of increased molt frequency. Based on results from this study and from previous investigations, it is estimated that P. interruptus males and females become sexually mature by ages 4 and 5 years, respectively, and that legai size is reached by 7 or 8 years of age. Juvenile P. interruptus activity patterns and foraging behavior were similar to those of adults, except that juvenile home ranges were proportionally smaller, and small juveniles were apparently not attracted to distant food. Small mollusks, abundant in Phyllospadix habitats, were the major food items. Size-dependent predation by fish and octopus apparently caused the considerable juvenile mortality observed at Bird Rock. Juveniles approaching 2 years of age gathered in mixed size-class aggregations by day and foraged beyond the grass beds at night. In autumn, these juveniles migrated to deeper habitats, coincident with new puerulus settlement in the Phyllospadix beds. Based on strong inferences from the results, it is proposed that size-dependent predation is the most important factor determining the !ife history strategy of juvenile P. interruptus. Life history tactics promoting rapid growth apparently function dually in reducing the period of high vulnerability to predation and decreasing the time required to reach sexual maturity. The Phyllospadix habitat is an excellent lobster nursery because it provides shelter from predators and possesses abundant food resources for sustaining optimum juvenile growth rates in shallow, warm water.

Notes on the induced maturation and spawning in four-month-old Penaeus monodon Fabricius by eyestalk ablation

The ablation technique consisted of making an incision across the eyeball to allow free flow of fluids while holding the prawn under water, squeezing the eyeball contents outwards, and pinching hard the eyestalk tissue. The cut area heals completely in about a week; no application of antibiotics is necessary. Spent spawners were tagged with thin brass rings (Rodriguez, 1976) around the unablated eyestalk for a separate experiment on rematuration. Two spawning yielding approximately 277,000 eggs were obtained three weeks after ablation, followed four days later by two more spawnings with 160,000 eggs; all four spawners weighed more than 100 g. With a hatching rate of 98% and 78% for the first and second batch, respectively, the spawnings produced viable nauplii. Water temperatures as low as 23 degree C due to a delayed cold spell in March depressed molting; weakened larvae had to be discharged at the mysis stage. Although ovarian development continued, no further spawnings were obtained due mainly to the onset of bacterial and fungal disease. Infection is initiated in injured portions of the exoskeleton, sometimes penetrating right through the muscles to the ovarian tissues. The non-flowthrough conditions and mussel meat feeding led to fouling of the culture water resulting in consecutive mortalities caused by disease. Female P.monodon held in maturation pens were ablated at the age of 15 months (Santiago, et al., 1976); they averaged only 16 g body weight after four months growth in ponds. In another experiment, pond-reared P.monodon females ranging from 50 to 80 g were ablated at approximately seven months (Aquacop, 1977). The present results show a minimum age of four months from postlarve that P.monodon is capable of ovarian development and spawning upon ablation. However, maturation is probably affected by size as well as age - the four-month old females weighed an average of 100 g in contrast to the smaller animals in the earlier experiments.

Cloning and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .