Chemical and isotopic composition of karstic lakes in Jamaica,West Indies

Lakes in the limestone region of Jamaica exhibit a range of chemical characteristics that reflect varying inputs from precipitation, surface runoff and groundwater, together with the subsequent evolution of the water within the limnic environment. Detailed spatial and temporal sampling was conducted on one lake, Wallywash Great Pond. Chemical data, together with D/H, ¹⁸O/¹⁶O, ¹³C/¹²C and ⁸⁷Sr/⁸⁶Sr ratios confirm that the karstic spring waters entering the lake evolve chemically through degassing, mixing with rainfall and runoff, biogenic decalcification (resulting mainly from bicarbonate assimilation by the high biomass of submerged macrophytes), and evaporation. Modern carbonate sedimentation in Wallywash Great Pond is largely of high-Mg calcite. This is consistent with Mg/Ca molar ratios >2 within much of the lake. However, aragonite forms on the adaxial leaf surfaces of Potamogeton spp. This may be explained either as a result of locally elevated Mg concentrations or a high degree of supersaturation favouring very rapid carbonate precipitation. Two small lakes to the north of Wallywash Great Pond show minor influence of the Na-Cl dominated coastal aquifer, suggesting that coastal lakes are sensitive to variations in the boundary between fresh and brackish groundwater caused by changes in climate or sea level. Their ¹³C/¹²C ratios are strongly influenced by biogenic CO₂ derived from plant respiration or decay.     

Isolation and identification of Acanthamoeba species related to amoebic encephalitis and nonpathogenic free‐living amoeba species from the rice field

Aims: Isolation and characterization of the clinically relevant amphizoic amoebas in vegetated farmlands, which may present a risk to farmers’ health. Methods and Results: Acanthamoeba species was isolated and characterized via morphological and molecular means in the rice field where the patient was exposed to rice paddy water which most probably was the point of infection. An Acanthamoeba sp. abundant in the rice field was identified. Genotyping showed the strain to be genotype T4, which was identical to the amoebic parasite found in patient’s cerebrospinal fluid. During the course of the study, three nonpathogenic free‐living amoeba species were also isolated and characterized for the first time in Taiwan. Conclusions: This study successfully located a possible source of granulomatous amoebic encephalitis in a patient and provided the first evidence that Acanthamoeba genotype T4 may be a potential pathogen in Taiwan. Significance and Impact of the Study: The integration of field survey, clinical data and morphological and genetic examination represents a sound strategy for investigation of the possible role of free‐living amoebae in causing human diseases. Future work should include investigating the potential contributory role of other nonpathogenic free‐living protozoa in disease of livestock or even human.     

Isolation and Molecular Characterization of Acanthamoeba Strains from Dental Units in Costa Rica

Free‐living amoebae are protozoa widely distributed in nature, which can be found in a variety of environments. Four genera are recognized as causal agents of infections in humans and animals: Acanthamoeba, Naegleria, Balamuthia, and Sappinia. In this study, the presence of Acanthamoeba in dental units was determined and the isolates obtained were molecularly characterized; osmotolerance and thermotolerance assays were also performed to evaluate multiplication under these conditions, frequently associated with pathogenicity. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of Acanthamoeba genotype T4 in 14% of the units sampled. Osmotolerance and thermotolerance tests were positive for more than 80% of the isolates. Up to date, this is the first study that reports the detection, identification, and genotyping of Acanthamoeba isolated from dental units in Costa Rica and even in Latin‐America. Further assays to determine the potential pathogenicity of these Acanthamoeba isolates are underway.     

Foraminiferal paleoecology of the Montpelier and lower coastal groups (eocene-miocene), Jamaica,West Indies

Two domains of carbonate deposition characterized mid-Tertiary Jamaica. After latest Cretaceous to Paleocene orogeny, submergence of insular paleo-Jamaica accompanied the strike-slip or extensional faulting associated with the formation of the Cayman Trench to the north. Differential subsidence along a series of peripheral subsea escarpments (Duanvale-Wagwater escarpment) produced relief in excess of 2000 m by the Late Eocene. Shoalwater limestones covered the slowly subsiding Cornwall-Middlesex platform, thus ending the supply of clastics to deep-sea bottoms north and east of the escarpment where contemporaneous planktonic-foraminiferal pelagites accumulated. Middle Eocene to Middle Miocene carbonate rocks deposited in the deep-sea represent a distinctive lithogenetic unit termed the Montpelier Group.A preponderance of globigerinacean and radiolarian tests characterizes lower Montpelier microfossil assemblages. Dominant benthonic forms include Melonis pompilioides, Fontbotia wuellerstorfi and species of Stilostomella and Pleurostomella. Available faunal criteria including the assemblage composition, depth preferences of extant species and recurrent morphologic-ecologic patterns suggest abyssal (2000 m) paleo-depths at the site of accumulation on a sediment apron near the base of the Duanvale-Wagwater escarpment. Computed from inferred paleodepth and estimated sedimentary thickness, Middle Eocene to Lower Miocene subsidence totals 2800 m. Biostratigraphic and paleoecologic data do not support the prevalent concept of a regional unconformity within the Montpelier.In the Middle Miocene, regional uplift led to the emergence of the Cornwall-Middlesex platform and to pronounced shoaling of marginal sea bottoms. Here, hemipelagic sedimentation resumed during the later Middle Miocene after the carbonate veneer on the adjacent platform was sufficiently eroded so as to expose noncarbonate rocks. See NAPS document No. 02395 for 67 pages of faunal reference lists, maps of sample locations and tables of foraminiferal occurrences. Order from ASIS/NAPS, c/o Microfiche Publications, 305 East 46th St., New York, N.Y., U.S.A. 10017. Remit with order $9.05 for photocopies or $5.55 for microfiche. Make checks payable to Microfiche Publications.     

中国五省市甲型肝炎病毒基因分型的研究

为了解甲型肝炎（甲肝）病毒（ＨＡＶ）在中国几个城市的基因型分布,选择浙江杭州、江苏启东、安徽铜陵、云南昆明和上海市等的甲肝病人粪便标本或血清标本,以逆转录－套式聚合酶链反应（ＲＴ－ｎＰＣＲ）扩增合成ＨＡＶ ＶＰ１／２Ａ交接区基因区,并进行直接核苷酸序列分析和差异比较。结果表明,从这些城市甲肝病人分离到的１７株ＨＡＶ株均属基因Ⅰ型,为ＩＡ和ＩＢ亚型;所有ＨＡＶ株间核苷酸差异均小于１５％,但约５０％Ｈ     

Reproduction, diet and population structure of the mountain mullet, Agonostomus monticola, in Jamaica, West Indies

Reproduction, feeding and population structure were investigated in the mountain mullet, Agonostomus monticola, in eastern Jamaica between 1982 and 1986. Female fishes just outnumbered males 1.2 : 1. Minimum size at maturity for females was 123 mm fork length (FL) and 96 mm FL for males. Mean size at maturity for females was 165.8 mm FL (S.D.=1.61) and for males was 148.4 mm FL (S.D.=2.97). Fecundity was 340 000 eggs for a 25 g ovary. Fecundity (F) was related to size by the equation F=153.8 FL^1.36. Mean lengths of males and females captured by all methods were 129.4 mm (S.D.=5.56) and 160.4 mm FL (S.D.=0.70), respectively. A. monticola was found to be chiefly an insectivorous feeder. The major food items found in stomach contents in ranked sequence were insects, freshwater prawns, plant detritus and algae.     

Isolation, tissue distribution, and molecular characterization of Toxoplasma gondii from chickens in Grenada, West Indies

The prevalence of Toxoplasma gondii in free-range chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 102 free-range chickens (Gallus domesticus) from Grenada was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT). Antibodies were found in 53 (52%) chickens with titers of 1:5 in 6, 1:10 in 4, 1:20 in 4, 1:40 in 4, 1:80 in 15, 1:160 in 9, 1: 320 in 5, 1:640 in 4, and 1:1,280 or greater in 2. Hearts, pectoral muscles, and brains of 43 seropositive chickens with MAT titers of 1:20 or greater were bioassayed individually in mice. Tissues of each of 10 chickens with titers of 1:5 and 1:10 were pooled and bioassayed in mice. Tissues from the remaining 49 seronegative chickens were pooled and fed to 4 T. gondii-free cats. Feces of cats were examined for oocysts; they did not shed oocysts. T. gondii was isolated from 35 of 43 chickens with MAT titers of 1:20 or greater; from the hearts, brains, and pectoral muscles of 2, hearts and brains of 20, from the hearts alone of 11, and brains alone of 2. T. gondii was isolated from 1 of 10 chickens with titers of 1:5 or 1:10. All 36 T. gondii isolates were avirulent for mice. Genotyping of these 36 isolates using polymorphisms at the SAG2 locus indicated that 29 were Type III, 5 were Type I, 1 was Type II, and 1 had both Type I and Type III. Genetically, the isolates from Grenada were different from those from the United States; Type II was the predominant type from the United States. Phenotypically, all isolates from Grenada were avirulent for mice, whereas those from Brazil were mouse-virulent. This is the first report of isolation of T. gondii from chickens from Grenada, West Indies.     

Isolation and characterization of a cDNA encoding a mammalian cathepsin L-like cysteine proteinase from Acanthamoeba healyi

We have cloned a cDNA encoding a cysteine proteinase of the Acanthamoeba healyi OC-3A strain isolated from the brain of a granulomatous amoebic encephalitis patient. A DNA probe for an A. healyi cDNA library screening was amplified by PCR using degenerate oligonucleotide primers designed on the basis of conserved amino acids franking the active sites of cysteine and asparagine residues that are conserved in the eukaryotic cysteine proteinases. Cysteine proteinase gene of A. healyi (AhCP1) was composed of 330 amino acids with signal sequence, a proposed pro-domain and a predicted active site made up of the catalytic residues. Cys25, His159, and Asn175. Deduced amino acid sequence analysis indicates that AhCP1 belong to ERFNIN subfamily of C1 peptidases. By Northern blot analysis, no direct correlation was observed between AhCP1 mRNA expression and virulence of Acanthamoeba, but the gene was expressed at higher level in amoebae isolated from soil than amoeba from clinical samples. These findings raise the possibility that Ahcp1 protein may play a role in protein metabolism and digestion of phagocytosed bacteria or host tissue debris rather than in invasion of amoebae into host tissue.     

Mutants of Acanthamoeba castellanii Resistant to Erythromycin, Chloramphenicol, and Oligomycin

SYNOPSIS. Cell lines of Acanthamoeba castellanii resistant to erythromycin (Ery^R), chloramphenicol (Cap^R), and oligomycin (Oli^R) have been isolated. These may be the first such mutants for A. castellanii. These mutants have been phenotypically stable for 2 years, surviving storage and vegetative multiplication in the absence of drugs. Resistance was specific for each drug, but double mutants (e.g. Ery^RCap^R) were obtained by stepwise selection. Mutant frequencies were determined in multiwell plates; <10 colony forming units (CFU/10⁵ amebas were observed in wild-type populations 12 days after incubation in 500 μg Ery/ml, 2.5 mg Cap/ml, or 15 μg Oli/ml. After 30 days, averages of 100 CFU/10⁵ amebas were observed in Ery and Cap, whereas, frequencies for Oli remained unchanged. Frequencies for Ery^R and Cap^R were consistent with rates of recovery from these drugs in batch cultures. We were unable to obtain spontaneous mutants resistant to cycloheximide, emetine, 5-fluorodeoxyuridine, or ethidium bromide. Ery^R, Cap^R and Oli^R could be mitochondrial mutants.     

Upper Eocene to Middle Miocene ostracode faunas and paleo-oceanography of the North Coastal Belt,Jamaica, West Indies

The ostracode fauna of the Montpelier Group (Upper Eocene-Middle Miocene) exposed in the western part of the North Coastal Belt of Jamaica contains representatives of three paleo-environments. Allochthonous shelf and littoral species, characterized by abraded carapaces and eye tubercles, and a diverse group of archibenthal (slope) forms are present throughout. Species restricted to the World Ocean Psychrosphere (> 1000 m), notably Bradleya dictyon, Australoecia tipica, Agrenocythere hazelae and Macrocypris spp., first occur in sediments of earliest Miocene age (planktonic foraminiferal zone N4). Their presence establishes a minimum age for the entry into the Cayman Trench of frigid (<8–10°C) water masses drawn from the Atlantic thermohaline stratification.Upper Eocene and Oligocene assemblages are dominated by smooth, blind genera such as Krithe, Messinella and Bythocypris and lived in lower thermospheric conditions (10°C) at or near the base of the thermocline (700– 900 m). During the Early Miocene, downfaulting of the north island slope of Jamaica in response to southward tilting of the Nicaraguan Rise brought the Montpelier depositional site into contact with progressively deeper and colder water-masses. This subsidence culminated in the late Early Miocene; faunas of this age contain large numbers of Bradleya dictyon, Agrenocythere hazelae, Macrocypris sp. 1, Bairdia oarion and Bairdoppilata cassida. They suggest paleodepths of 1500–2000 m and the local presence of a 3–4° water-mass tentatively identified as the North Atlantic deep water. If this interpretation is correct, then the maximum sill-depth and water-mass configuration of the Miocene Cayman Trench have persisted to the present. Middle Miocene assemblages indicate a return to paleodepths of 1000–1500 m heralding the pronounced Upper Miocene to mid-Pleistocene tectonism which elevated Montpelier strata above sea level. An alternative view attributes the Miocene ostracode succession in the Montpelier to episodes in the development of the Atlantic circulation as recognized in the northeast Atlantic.Montpelier deposition took place either on a broad step intervening between the northern edge of the Clarendon Platform and the abyssal basins of the Cayman Trench or on a Tertiary analog of the existing north island slope. These results have emphasized the value of detailed taxonomic and morphologic studies of deep-sea ostracodes in geohistorical reconstruction.     

Light and Electron Microscopic Observations on Encystment of Acanthamoeba palestinensis, Reich

SYNOPSIS. The ultrastructural changes occurring during encystment of Acanthamoeba palestinensis have been investigated. The cyst wall consists of endocyst and exocyst, both having the same fine structure. At irregular intervals in the cyst wall ostioles occupied by opercula are present. The nuclear membrane forms bulb-shaped projections and releases vesicles bounded by double membranes into the cytoplasm. Dense nucleolus-like bodies of different sizes and variable numbers are found in the nucleus of every cyst. The importance of the cyst structure as a taxonomic criterion is discussed.     

鸡肠源芽胞杆菌的分离、鉴定和抗菌活性

从人工饲养的健康三黄肉鸡的盲肠和大肠中分离出10株芽胞杆菌,对其进行菌落形态观察、革兰染色、芽胞染色和生理生化特性鉴定,确定为枯草芽胞杆菌、地衣芽胞杆菌、蜡状芽胞杆菌、巨大芽胞杆菌、球形芽胞杆菌、短小芽胞杆菌、凝结芽胞杆菌。同时测定了它们对动物病原性大肠埃希菌和链球菌的抑菌活性。其中有9株芽胞杆菌对大肠埃希菌有抑制作用,8株芽胞杆菌对链球菌有抑制作用。结果表明芽胞杆菌分离株Y2、Y3、Y4、Y5、Y6、Y7、Y8具有作为益生菌开发的潜力。     

In-vitro activity of miltefosine and voriconazole on clinical isolates of free-living amebas: Balamuthia mandrillaris, Acanthamoeba spp., and Naegleria fowleri

The anticancer agent miltefosine and the antifungal drug voriconazole were tested in vitro against Balamuthia mandrillaris, Acanthamoeba spp., and Naegleria fowleri. All three amebas are etiologic agents of chronic (Balamuthia, Acanthamoeba) or fulminant (Naegleria) encephalitides in humans and animals and, in the case of Acanthamoeba, amebic keratitis. Balamuthia exposed to <40 microm concentrations of miltefosine survived, while concentrations of >or=40 microM were generally amebacidal, with variation in sensitivity between strains. At amebastatic drug concentrations, recovery from drug effects could take as long as 2 weeks. Acanthamoeba spp. recovered from exposure to 40 microM, but not 80 microM miltefosin. Attempts to define more narrowly the minimal inhibitory (MIC) and minimal amebacidal concentrations (MAC) for Balamuthia and Acanthamoeba were difficult due to persistence of non-proliferating trophic amebas in the medium. For N. fowleri, 40 and 55 microM were the MIC and MAC, respectively, with no trophic amebas seen at the MAC. Voriconazole had little or no inhibitory effect on Balamuthia at concentrations up to 40 microg/ml, but had a strong inhibitory effect upon Acanthamoeba spp. and N. fowleri at all drug concentrations through 40 microg/ml. Following transfer to drug-free medium, Acanthamoeba polyphaga recovered within a period of 2 weeks; N. fowleri amebas recovered from exposure to 1 microg/ml, but not from higher concentrations. All testing was done on trophic amebas; drug sensitivities of cysts were not examined. Miltefosine and voriconazole are potentially useful drugs for treatment of free-living amebic infections, though sensitivities differ between genera, species, and strains.     

Molecular Characterization of Acanthamoeba spp. Occurring in Water Bodies and Patients in Poland and Redefinition of Polish T16 Genotype

Acanthamoeba genus is divided into 20 genotypes (T1–T20) on the basis of the gene encoding 18S rRNA sequence. Using of at least 2 kbp gene fragments is strongly recommended to identify new genotypes and 5% difference is commonly used as a criterion of new genotypes, however, this value is questionable. In this paper, Polish Acanthamoeba strains described earlier on the basis of ~850 bp Ami fragment of 18S rRNA gene as T4, T11 and a new T16 genotype, have been analyzed using near‐complete sequence of the gene. This analysis was needed because the Ami fragment does not reveal full variability within 18S rRNA gene. Phylogenetic analysis based on Ami fragment is biased by artifacts in the construction of the tree, so the fragment should not be used for identification of new putative Acanthamoeba genotypes. The analysis confirmed that the Polish sequences represent T4 and T11 genotypes and that the strains described earlier as T16 genotype are in fact a new subgroup of the T20 genotype and that this genotype should be divided into two subgroups: T20a (two strains described by [J. Eukaryot. Microbiol. 62 (2015) 69]) and T20b (11 Polish strains described in this study). The T20b subgroup was isolated from both clinical samples and water bodies used by people as bathing places and there is a risk of infection for humans during contact with water.     

抗动物病原菌芽孢杆菌的筛选、初步鉴定和抗菌活性

从鸡肠道分离、挑选的18株芽孢杆菌经培养特征、形态观察、生理生化实验,初步被鉴定为枯草芽孢杆菌(Bacillus subtilus)、地衣芽孢杆菌(Bacillus licheniformis)、蜡状芽孢杆菌(Bacillus cereus)、巨大芽孢杆菌(Bacillus megaterium)和凝结芽孢杆菌(Bacillus coagulans)。同时测定了它们对5种常见动物病原菌大肠杆菌(E.coli)、金黄色葡萄球菌(Staphylococcus aureus)、肠炎沙门氏菌(Salmonella typhimurium)、猪链球菌(Streptococcus suis)、奇异变形杆菌(Proteus mirabilis)的抑菌活性,其中有4株芽孢杆菌对5种病原菌都有抑制作用。还分别测定了它们产木聚糖酶(从0.895 U1到3.239 U1)和纤维素酶活性(分别为0.391 U2和0.465 U2)。结果表明,芽孢杆菌分离株BC17、BC32、BC106、BC228、BC247和BC261具有作为益生菌的开发潜力。     

Osteology of a slave burial population from Barbados,West Indies

A unique seventeenth–nineteenth century slave cemetery population from Newton plantation, Barbados, allows examination of craniodental characters in relation to ethnohistorical data. Age-at-death estimates suggest life expectancy at birth of 29 years and low infant mortality; historical demography, however, suggests life expectancy of 20 years and very high infant mortality. Tooth decay, bilateral tooth loss, periodontal disease, root hypercementosis, and severe enamel hypoplasia are high in frequency. The teeth yield evidence of such cultural practices as pipe-smoking and incisor mutilation. Several skeletal features reflect periodic near-starvation. Directional and fluctuating dental asymmetry, relative tooth size, and hypoplasia distribution suggest slaves experienced considerable weaning trauma; metabolic stress at this time exceeded that of prenatal and immediate postnatal periods. Odontometrics and dental and cranial nonmetric traits indicate that modern Blacks are intermediate between the ancestral slaves and modern Whites but more similar to the latter, suggesting effects of environmental covariance exceed those of genetic admixture. Nonmetric trait distributions show nonrandom patterns according to area of burial in the cemetery, a possible result of family segregation.     

Isolation and characterization of probiotic properties of Lactobacilli isolated from rat fecal microbiota

The objective of the present study was to characterize lactobacilli isolates from the feces of male Wistar rats. Various physiological features of the candidate probiotic isolates were preliminarily investigated, including tolerance to simulated gastric juice and bile salts, antimicrobial activity, antibiotic susceptibility and in vitro aggregation. Based on their morphological and biochemical characteristics, four potential probiotic isolates (CS2, CS3, CS4, and CS7) were screened. The isolates showed good tolerance to stimulated gastric juice and bile salts. CS4 and CS7 exhibited strong antibacterial activities against the pathogens tested as assessed in neutral pH culture supernatants. All lactobacilli isolates were susceptible to all the tested antibiotics, except vancomycin. Moreover, the isolate CS4 and CS7 were found to possess stronger cell surface traits such as hydrophobicity, auto‐aggregation and co‐aggregation capacity. In addition, CS4 and CS7 had greater β‐galactosidase activities than the others. Biochemical tests and 16S rRNA gene sequencing confirmed that CS2, CS3, CS4 and CS7 are Lactobacillus intestinalis PJ2, L. sakei PJ3, L. helveticus PJ4, and L. plantarum PJ7, respectively. Based on the obtained results, L. helveticus PJ4 and L. plantarum PJ7 are ideal in vitro probiotic candidates and require further in vivo evaluation.     

凡纳滨对虾源不动杆菌群体感应信号分子分离鉴定及其调控

【目的】鉴定凡纳滨对虾源不动杆菌(Acinetobacter spp.M1)分泌的N-酰基高丝氨酸内酯(AHLs)类型,探究细菌生长阶段及环境因素对其分泌信号分子的影响。【方法】报告菌株平板法检测M1的AHLs的活性;采用报告平板与薄层层析(TLC)相结合法对M1分泌的AHLs类型进行鉴定。【结果】菌株M1分泌N-3-氧代-己酰基-高丝氨酸内酯和N-3-氧代-辛酰基-高丝氨酸内酯两种信号分子。在适宜条件下AHLs活性随着培养时间的延长先升高后降低,在对数末期(30 h)达到最大。弱酸和弱碱环境能够降低M1分泌AHLs的能力,p H 7.0是M1分泌AHLs的最适p H。较高浓度的Na Cl促进了个体M1分泌AHLs的能力,但是Na Cl浓度对M1总体分泌AHLs没有显著的影响。菌株M1分泌AHLs的最佳温度为30°C,温度过高或过低都会影响其分泌。【结论】菌株M1主要产生N-3-氧代-己酰基-高丝氨酸内酯和N-3-氧代-辛酰基-高丝氨酸内酯两种类型信号分子。M1的QS系统受菌体密度和环境因素的双重调控。     

Isolation and characterization of diazotrophic bacteria from banana and pineapple plants

Banana and pineapple fruit crops are widely cultivated in tropical areas where high amounts of fertilizers are applied, principally nitrogen. Over 200 kg N.ha^-1.yr^-1 is often applied to these crops. Nevertheless, developing countries face the problem of high costs of chemical fertilizers. As already demonstrated for other tropical crops, like sugar cane, the utilization of nitrogen-fixing bacteria may support the growth of these fruit plants. In this work, we demonstrate the association of nitrogen-fixing bacteria with banana and pineapple. Samples from roots, stems, leaves and fruits of different genotypes showed the occurrence of diazotrophic bacteria, when evaluated in semi-specific semi-solid media. These isolates could be separated into seven different groups according to their morphological and physiological characteristics. Additional, phylogenetic assignments were performed with group- and species-specific oligonucleotide probes. Bacteria related to the groups of Azospirillum amazonense, Azospirillum lipoferum, Burkholderia sp. and a group similar to the genus Herbaspirillum could be detected in samples of both crops. However, Azospirillum brasilense and another two groups of Herbaspirillum-like bacteria were detected only in banana plants. Two isolates of the latter group were identified as Herbaspirillum seropedicae, whereas the other isolates may represent a new Herbaspirillum species.