Assessment of the in vivo genotoxicity of a new formulation of Bacillus thuringiensis var israelensis SH-14

Biological control agents have become a useful alternative for the reduction of the use of chemical insecticides. LABIOFAM (Cuba) is developing a new formulation of a biolarvicide that possesses as active biological agent Bacillus thuringiensis var israelensis serotype H14. In order to evaluate the genotoxicity of this new formulation, an in vivo battery test was used: micronucleus (MN), chromosome aberrations (CAs), and sperm morphology (SM) assays. A dose of 6.45?×?10⁸ spores was administered per animal via oral administration. Bone marrow cells were collected 24?h after a two day treatment for the MN assay, and 24?h after a unique treatment for the CA assay, using cyclophosphamide as the positive control. Sperm cells were collected at 5 weeks from the first of five administrations for the SM test, using acrylamide as positive control. Bacillus thuringiensis var israelensis serotype H14 failed to show either a significative increase of micronucleated polychromatic erythrocytes, chromosomal aberrations, or sperm abnormalities. Acute oral administration of a high dose of Bacillus thuringiensis var israelensis serotype H14 did not produce mutagenic effects in bone marrow or sperm cells.     

Leaf ontogeny interacts with Bt modification to affect innate resistance in GM aspens

Bioassays with a non-target slug (Deroceras spp.) and chemical analyses were conducted using leaf tissue from already existing genetically modified insect-resistant aspen trees to examine whether genetic modifications to produce Bacillus thuringiensis (Bt) toxins could affect plant phytochemistry, which in turn might influence plant–herbivore interactions. Three major patterns emerged. First, two independent modifications for Bt resistance affected the phytochemical profiles of leaves such that both were different from the isogenic wild-type (Wt) control leaves, but also different from each other. Among the contributors to these differences are substances with a presumed involvement in resistance, such as salicortin and soluble condensed tannins. Second, bioassays with one Bt line suggest that the modification somehow affected innate resistance (“Innate” is used here in opposition to the “acquired” Bt resistance) in ways such that slugs preferred Bt over Wt leaves. Third, the preference test suggests that the innate resistance in Bt relative to Wt plants may not be uniformly expressed throughout the whole plant and that leaf ontogeny interacts with the modification to affect resistance. This was manifested through an ontogenetic determined increase in leaf consumption that was more than four times higher in Bt compared to Wt leaves. Our result are of principal importance, as these indicate that genetic modifications can affect innate resistance and thus non-target herbivores in ways that may have commercial and/or environmental consequences. The finding of a modification–ontogeny interaction effect on innate resistance may be especially important in assessments of GM plants with a long lifespan such as trees.     

Deponiegas

Investigations were performed concerning whether or not the composition of volatile halogenated carbons in landfill gas (LFG) change when they uncontrolledly leave the aerobic surface of landfill. For this reason, the aerobic biological degradation of chlorofluorocarbons (CFCs), chlorofluorohydrocarbons (HCFCs), and vinyl chloride (VC), which were often measured in LFG of old household-landfills, was investigated. In 1-liter and 126-liter laboratory test reactors, the aerobic milieu conditions of the surface of landfills were simulated. Air with methane and trichlorofluoromethane (R11), dichlorodifluoromethane (R12), and their reductive metabolites dichlorofluoromethane (R21), chlorodifluoromethane (R22), and vinyl chloride (VC) were continuosly pumped through marl and a mixture of biowaste and compost. R21, R22, and VC were degraded, probably cometabolically by methanotrophic bacteria. They have inhibited the methane oxidation. The degradation rates of R21 (only in biowaste and compost) and R22 were up to 2.8 and 10 mg/m³ _material/h, respectively. The maximum degradation rate of VC was 1,3000 mg/m³ _material/h in marl. Under aerobic milieu conditions, a biodegradation of R11 and R12 was not detected.     

Differential toxic response of Chironomus travalensis kieffer ( Chironomidae : Diptera ) to some industrial effluents

Acute toxicity testing of some industrial effluents (rubber (Rr), brewery (Br), and bottling (Bt)) on Chironomus travalensis (Diptera larvae) was determined at 96-h using a static bioassay technique. Effluent quality characterization revealed acidic pH values of 4.5 for Rr, 5.64 for Br, and 4.32 for Bt effluents. Tests were carried out in replicates between 10 and 100% effluent concentrations. Mortality rate rose progressively with increasing concentrations. Bt effluent was the most toxic with 96-h LC₅₀ of 61.5%, closely followed by Rr effluent with 96-h LC₅₀ of 87% and the least toxic was Br effluent with 96-h LC₅₀ of 90%.     

Detection of Escherichia coli in wastewater based on enzyme immunoassay

This research describes a fast detection method on the basis of enzyme-linked immunosorbent assay (ELISA) for Escherichia coli in drainage of wastewater treatment plants. Optimized conditions such as the reaction format (sandwich or direct), the concentrations of diluted horseradish peroxidase (HRP)-E. coli conjugate, and anti-HPR antibody and pretreatment of E. coli were studied. Those results showed that the linear range of detection for E. coli was 10 cfu/mL-6 × 10⁴ cfu/mL. Compared with conventional methods, it is a convenient and sensitive detection method with low cost.     

双酚A对小鼠肝脏和肾脏细胞的氧化损伤

为探究双酚A(BPA)的氧化毒性,分别以剂量为20、40和80mg·kg~(-1)·d~(-1)的BPA对雄性昆明小鼠灌胃处理1周,并测定了小鼠体内活性氧自由基(ROS)水平、还原型谷胱甘肽(GSH)含量、丙二醛(MDA)含量和DNA-蛋白质交联系数(DPC)。与对照组相比,各BPA暴露组小鼠肝脏和肾脏细胞中的ROS生成量、MDA含量和DPC系数均升高,而GSH含量下降(P<0.05或P<0.01)。ROS生成量、GSH含量和DPC系数均显示出剂量-效应关系。研究表明,BPA可扰乱小鼠肝脏和肾脏细胞的氧化应激平衡,诱导细胞氧化损伤。     

盐酸小檗碱对小鼠脾细胞的DNA损伤和氧化性损伤研究

为了探讨盐酸小檗碱对小鼠的DNA损伤和氧化性损伤。随机选取30只小鼠分成对照组以及7.5,15,30,60与120 mg?kg^-1实验组,处理后,应用小鼠脾细胞进行彗星实验与抗氧化酶实验。测定DNA损伤情况以及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、过氧化物酶（POD）活性以及丙二醛（MDA）含量变化。对盐酸小檗碱的DNA损伤与氧化性损伤作用进行比较研究。研究结果表明：彗星实验中,随着盐酸小檗碱浓度的增加,尾部DNA含量、尾长与尾矩均增加,与阴性对照组相比差异有统计学意义（p<0.05或p<0.01）,且呈剂量-效应关系;超氧化物歧化酶(SOD)与过氧化氢酶(CAT)活性随盐酸小檗碱剂量增加逐渐降低,丙二醛(MDA)含量明显下降,过氧化物酶(POD)活性在7.5 mg?kg^-1时上升,而后逐渐下降。在60 mg?kg^-1和120 mg?kg^-1时,有极显著性差异（p<0.01）产生。由此可见,盐酸小檗碱对小鼠脾细胞有一定的损伤作用,能够引起小鼠脾细胞的DNA损伤和氧化性损伤。     

Labyrinthula sp., a marine slime mold producing the symptoms of wasting disease in eelgrass,Zostera marina

Coastal ecosystems along the eastern United States are presently threatened by a recurrence of the wasting disease of eelgrass, Zostera marina L. Using Koch's postulates, a species of the marine slime mold, Labyrinthula, is identified as the causal microorganism of this disease. Our disease tests for pathogenicity performed on eelgrass, using four Labyrinthula spp., indicate only one species produces the disease symptoms identical to those found associated with the wasting disease. The pathogenic Labyrinthula sp. has morphological characteristics that distinguish it from the other three species. Identification of Labyrinthula spp. is difficult because species described in the literature are not clearly characterized or identifiable. Tests at various salinities demonstrate that disease symptoms appear infrequently at salinities of 10%. or less.     

Ultrastructure of the vitelline coat in the ascidians Phallusia mammillata, Ascidia mentula and Ciona intestinalis : new aspects revealed by freeze-substitution and deep-etching

The microstructure of the vitelline coat (VC) of the three ascidian species Phallusia mammillata, Ascidia mentula and Ciona intestinalis was analyzed by transmission electron microscopy (TEM) using freeze-substitution and deep-etching. In addition, eggs were examined by light microscopy and prepared by conventional chemical fixation methods for TEM in order to illustrate the advantages of cryopreparation for the preservation of the VC ultrastructure. In conventionally fixed eggs of P. mammillata and A. mentula the VC appeared as a tripartite structure, consisting of a thin outer fibrous layer (OFL), a central dense layer (CDL) and a relatively wide inner fibrous layer (IFL). In C. intestinalis a sparse OFL is made up by fine tufts of filaments but an IFL is lacking. In all three species the test cells were located in the area between the inner VC boundary and the egg cell membrane. Using our modified freeze-substitution method a clear improvement in preservation of the VC matrix was obtained. In all three species the OFL as well as the IFL in P. mammillata and A. mentula appeared as extended fibrous meshworks. Furthermore, the area of the perivitelline space, empty after conventional TEM preparation, was found to be filled by an additional matrix, designated here as perivitelline fibrous matrix (PVFM). Quick-freeze/deep-etch/rotary-shadowing enabled us to confirm the overall architecture of the egg matrix and allowed a further detailed investigation of the VC fine structure in P. mammillata. Accordingly, the vitelline coat of P. mammillata and A. mentula is a multilayered matrix composed of four distinct subunits (OFL, CDL, IFL, PVFM), whereas in C. intestinalis an IFL is lacking. Received: 7 November 1996 / Accepted: 5 December 1996     

不同补给水源的城市人工湖中异养菌耐药性及耐药菌种属分布研究

为了阐明不同补给水源的城市人工湖中异养菌耐药状况和耐药菌种属分布特征,选取分别以地表水和再生水为补水来源的XQ湖和FQ湖为代表进行研究。从2018年4—11月逐月采集水样,考察了各水样中对氨苄西林(AMP)、磺胺甲恶唑(SMZ)和四环素(TET)这3种不同种类抗生素具有耐药性的异养菌含量,并对分离菌株的耐药表型、耐药菌株的种属分布以及水质理化指标进行了分析。结果表明,2处人工湖中的AMP耐药菌和SMZ耐药菌含量约为10~2～10~4CFU·(100 m L)~(-1),而TET耐药菌含量则约10~1～10~3CFU·(100 m L)~(-1)。分离出的84株耐药菌归属于19个种,其中蜡状芽孢杆菌、大肠埃希氏菌、维氏气单胞菌、豚鼠气单胞菌和鸟氨酸拉乌尔菌为2个湖共有耐药菌。71.4%的耐药菌都是对AMP单一耐药,以蜡状芽孢杆菌为主。由于具有固有耐药性的细菌在分离出的耐药菌中占比很低,获得性耐药很可能在城市人工湖中异养菌耐药性的发展上发挥了主要贡献作用。地表水补水的XQ湖和再生水补水的FQ湖在总异养菌含量、耐药菌含量和检出率上均无显著差异。分离来源对16株耐药性气单胞菌的聚类无明显影响。     

Resting spore formation of the marine planktonic diatom Chaetoceros anastomosans induced by high salinity and nitrogen depletion

The marine planktonic diatom Chaetoceros anastomosans, which was isolated from Sagami Bay, was used for a study of resting spore formation mechanisms in batch culture experiments. Vegetative cells could grow at salinities ranging from 20.7 to 45.5‰, and resting spore formation was enhanced significantly in nitrate-depleted, high salinity media (40.0 to 45.5‰). The rate of resting spore formation (1.9 d⁻¹) was comparable to the specific growth rate (1.8 d⁻¹) of vegetative cells in the exponential growth phase in normal salinity medium. The size of resting spores formed under high salinity conditions was smaller than that of spores formed in normal salinity media. Unlike vegetative cells, resting spores seemed to possess some mechanisms to survive over a wider range of salinities by resisting bacterial attacks on their cell walls. Received: 4 August 1996 / Accepted: 27 August 1996     

Tertiary-butyl acetate metabolism,toxicity, and human health considerations

Tertiary-butyl acetate (TBAC) (CAS No. 540-88-5) is an organic solvent with a potential for occupational and environmental exposure as a result of its use in industrial coatings, adhesives, inks, and degreasers. The objective of these studies was to extend the toxicological database upon which health hazard and risk assessments of TBAC can be made. The metabolism of TBAC was studied in rats exposed by inhalation for 6?h to concentrations of 100 or 1000?ppm. There was an evidence of partial saturation of TBAC absorption and metabolism at some concentration below 1000?ppm. Approximately 5% of the low dose and 26% of the high dose was expired without change within 12?h, while the retained material was rapidly metabolised and excreted, mostly in the urine, within 24?h. Very little radioactivity remained in the tissues after day 7. The metabolism of TBAC appears to follow two major routes: hydroxylation of the tertiary-butyl moiety to form 2-hydroxymethylisopropyl acetate and ester hydrolysis to form tertiary-butyl alcohol (TBA). A minor route involves oxidation of the acetate moiety. Based on the proportion of metabolites that can clearly be assigned to one or the other major pathway, hydroxylation of the tertiary-butyl moiety prevails at 100?ppm, while hydrolysis of the ester bond predominates at 1000?ppm.

Based on nose-only inhalation exposure of rats to TBAC for 6?h, the LC50 for males and females combined is approximately 4200?ppm. Clinical signs included exaggerated breathing, staggering, tremors, and lethargy approximately 1?h after the exposure, but all surviving rats appeared normal from 24?h until the end of the 14?day observation period. An LC₅₀ was not identified for mice. After exposure of whole body for 6?h to 3000?ppm, the highest concentration tested, all mice were prostrate for most of the exposure time, but there were no deaths.

Groups of five male and five female Sprague-Dawley rats were exposed nose-only to 0, 100, 400, or 1600?ppm TBAC 6?h?day^?1 5 days?wk^?1 for 2?weeks. There were no effects on body weight, feed or water consumption, or necropsy findings. Male rats exposed to 1600?ppm had increased liver weights and hypertrophy of centrilobular hepatocytes. An increased proportion of cortical tubule cells with hyaline droplet accumulation was observed in all treated groups of males. Groups of five male and five female CD-1 mice were exposed whole body to 0, 190, 375, 750, or 1500?ppm TBAC 6?h?day^?1 for 14 consecutive days. There were no effects on body weight, feed consumption, or necropsy findings. Liver weights were increased in female mice at 750 and 1500?ppm. Minimal hypertrophy of centrilobular hepatocytes was found in female mice at 375, 750, and 1500?ppm and in male mice at 1500?ppm TBAC.

TBAC did not induce gene mutations in bacterial tests with strains of S. typhimurium and E. coli. Further, there was no evidence of clastogenic activity from tests either for the induction of chromosomal aberrations in human lymphocytes in vitro in the presence or absence of S9 mix or for the induction of micronuclei in bone marrow cells of rats exposed by inhalation for 6?h to 1600?ppm TBAC.

These results are relevant to human health risk assessment and are discussed in the context of previous studies. The weight of the scientific evidence supports the conclusion that TBAC has lower acute toxicity than previously suggested, that it is rapidly excreted when inhaled, and that neither TBAC nor its TBA metabolite are genotoxic or potential human carcinogens.     

As(V)-reduction to As(III) by arsenic-resistant Bacillus spp. bacterial strains isolated from low-contaminated sediments of the Oliveri-Tindari Lagoon,Italy

Five arsenic-resistant bacterial strains designated MT1, MT2, MT3, V1 and V2 were isolated from sediments of the Oliveri-Tindari Lagoon (Italy), which comprises six small lakes whose sediments contain low arsenic concentrations. Phylogenetic analysis of the 16S rRNA gene sequences assigned them to the genus Bacillus. Bacillus sp. strain MT3 showed higher tolerance to As(III) and As(V), as indicated by minimum inhibitory concentrations of 14 and 135 mmol^?1, respectively. Bacillus sp. strain V1 showed growth inhibition at 14 mmol^?1 in the presence of As(III) and at 68 mmol^?1 in the presence of As(V), whereas the arsenic resistance of Bacillus sp. strain MT1 was 10 and 27 mmol^?1 for As(III) and As(V), respectively. The strains Bacillus spp. MT2 and V2 showed low levels of As(III) and As(V) resistance, as it was unable to grow at concentrations>7 and 14 mmol^?1, respectively. The isolated arsenic-resistant Bacillus spp. strains were able to reduce As(V) to As(III), especially Bacillus spp. strain MT3. This study suggests that the isolated bacterial strains play a role in the arsenic biogeochemical cycle of arsenic-poor sediments in the Oliveri-Tindari Lagoon.     

Distribution,source apportionment and health risk assessment of polycyclic aromatic hydrocarbons (PAHs) in intertidal sediment of Asaluyeh,Persian Gulf

Surface sediment samples were collected from intertidal zone of Asaluyeh, Persian Gulf, to investigate distribution, sources and health risk of sixteen polycyclic aromatic hydrocarbons (PAHs). Total PAH concentrations ranged from 1.8 to 81.2 μg kg^?1 dry weight, which can be categorized as low level of pollution. Qualitative and quantitative assessments showed that PAHs originated from both petrogenic and pyrogenic sources with slight pyrogenic dominance. Source apportionment using principal component analysis indicated that the main sources of PAHs were fossil fuel combustion (33.59%), traffic-related PAHs (32.77%), biomass and coal combustion (18.54%) and petrogenic PAHs (9.31%). According to the results from the sediment quality guidelines, mean effects range-median quotient (M-ERM-Q) and benzo[a]pyrene toxic equivalents (BaP_eq), low negative ecological risks related to PAH compounds would occur in the intertidal zone of Asaluyeh. The total benzo[a]pyrene (BaP) toxic equivalent quotient (TEQ^carc) for carcinogenic compounds ranged from 0.01 to 7 μg kg^?1-BaP_eq, indicating low carcinogenic risk. The human health risk assessment of PAH compounds via ingestion and dermal pathways suggests low and moderate potential risk to human health, respectively.     

Disease resistance: a benefit of sociality in the dampwood termite Zootermopsis angusticollis (Isoptera: Termopsidae)

The benefit of sociality in relation to disease susceptibility was studied in the dampwood termite Zootermopsis angusticollis. Although contact with high concentrations of fungal conidia is lethal, the survivorship of nymphs exposed to spore suspensions ranging from 6 × 10⁶ to 2 × 10⁸ spores/ml of the fungus Metarhizium anisopliae increased with group size. The survivorship (measured as LT₅₀) of isolated individuals ranged from 3.0 to 4.8 days, but infected nymphs living in groups of 10 and 25 individuals survived significantly longer (5.6–8.3 and 5.6–9.1 days, respectively). In most cases, there were no significant differences in the survival distributions of the 10- and 25-termite groups. When nymphs were infected with concentrations of 7 × 10¹–7 × 10⁴ spores/ml and allowed to interact with healthy nestmates, fungal infections were not contracted by the unexposed termites. Moreover, infected termites benefitted from social contact with unexposed nestmates: their survival rates were significantly higher than those of infected termites living with similarly infected nestmates. Allogrooming, which increased in frequency during and after exposure to conidia, appeared to remove potentially infectious spores from the cuticle, thus increasing termite survivorship. These results suggest that allogrooming plays a crucial role in the control of disease and its death hazard in termites. The infection-reducing advantage of group living may have been significant in the evolution of social behavior in the Isoptera. Received: 18 March 1998 / Accepted after revision: 31 May 1998     

Chemopreventive role of Indigofera aspalathoides against 20-methylcholanthrene-induced carcinogenesis in mouse

This investigation was undertaken to evaluate the chemopreventive effect of hydroalcoholic extract of Indigofera aspalathoides (HAIA) against 20-methylcholanthrene (20-MC)-induced carcinogenesis in mice. Tumor was induced by single subcutaneous administration of 20-MC (200 µg per mouse) in Swiss albino mice. After 24 h of 20-MC administration, HAIA was administered at the doses of 250 and 500 mg kg^?1 body weight orally for 90 consecutive days. Mice of all groups were observed for 15 weeks to record tumor incidence (fibrosarcoma) and survival time. After 15 weeks the mice were sacrificed for the estimation of hematological profiles like hemoglobin (Hb), white blood cell (WBC), red blood cell (RBC), and liver biochemical parameters, namely lipid peroxidation, reduced glutathione (GSH), glutathione-S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT). HAIA treatment markedly reduced tumor incidence and prolonged the life span of sarcoma-bearing mice as compared to 20-MC control mice. Hematological profiles were significantly (p < 0.001) restored to normal levels in HAIA-treated mice as compared to 20-MC control mice. HAIA treatment significantly (p < 0.001) modulated the aforesaid liver biochemical parameters as compared to 20-MC control. The results concluded that I. aspalathoides demonstrated a remarkable chemopreventive effect in chemical-induced carcinogenesis in mouse. The potential chemopreventive action may be due to its antioxidant and detoxifying properties.     

Concentrations of polycyclic aromatic hydrocarbons in soils of a mangrove forest affected by forest fire

Surface soils affected by forest fires from Igbanko mangrove forest in Nigeria were analyzed for 16 EPA priority polycyclic aromatic hydrocarbons (PAHs) using gas chromatography–mass spectrometry (GC–MS). The total PAHs concentrations in the soils ranged from 63 to 188?µg?kg^?1 dry weight (average: 108?µg?kg^?1). The three predominant PAHs in the soils were naphthalene (Na), fluoranthene (Flu), and benzo(b)fluoranthene (BbF). Compared to the control sample (19?µg?kg^?1), elevated PAHs concentrations were observed in the soils, an indication of some level of PAHs contamination. PAHs source diagnostic ratios of Flu/(Flu?+?Pyr) and Ant/(Ant?+?Phe) indicated that the PAHs have a pyrogenic origin which may have resulted from combustion of grass, wood, or coal. An assessment based on Canadian soil quality guidelines indicated that the studied locations do not pose any serious adverse risk on human health.     

Resting spore formation and phosphorus composition of the marine diatom Chaetoceros pseudocurvisetus under various nutrient conditions

The marine diatom Chaetoceros pseudocurvisetus was used for a study of resting spore formation and cellular phosphorus composition. Resting spores were found in any culture medium with ample silica, including nitrogen limited, phosphorus limited and nutrient replete conditions. Resting spores protected themselves with thick silica walls, so that vegetative cells required about 3 pmol cell^-1 of additional silica to form resting spores. Phosphorus compounds in the cells were divided into eight fractions: nucleotide-P, orthophosphate, acid soluble polyphosphate, sugar phosphate, nucleic acid-P, acid insoluble polyphosphate, lipid-P and residual-P. The sum of orthophosphate, sugar phosphate and nucleic acid-P comprised over 65% of the total phosphorus in cells under any culture conditions. Sugar phosphate was the most variable component, being most abundant in vegetative cells and least abundant in resting spores.     

Clearance rates of the salp Thalia democratica fed naturally occurring particles

Net-captured aggregate stages of the salp Thalia democratica were fed naturally occurring particles in 2-liter bottles onboard ship. The goal was to compare clearance rates of net-captured T. democratica fed natural particles to clearance rates previously determined in laboratoryborn T. democratica fed monocultures of phytoplankton. On a cruise from 19–29 April, 1979, clearance rates were estimated by monitoring the decrease in concentration of particles with a multi-channel Coulter Counter TA II over a particle size range of 2 to 32 m equivalent spherical diameter (E.S.D.). Clearance rates were slightly lower than those measured in the laboratory, decreasing 3-fold over an 18-fold increase in the initial food concentration. Ingestion rates were slightly higher than those measured in the laboratory, increasing about 6-fold over the same range of food concentration. Within the particle concentration range examined (0.08 to 1.34 mm³l^-1), there was no lower threshold of food concentration below which clearance of particles ceased. There was no effect of the concentration of T. democratica on clearance rate over a range of 2.5 to 11.5 salps l^-1. The mean hourly ration was 2.0% of body carbon.     

Antioxidant rich diet supplements (Spirulina and tamarind fruit pulp) mitigate hematological disorders in fluoride exposed mice

The effect of antioxidant rich diet supplements of Spirulina and tamarind fruit pulp at 230 mg kg^?1 body weight, separately as well as in combination, on various hematological parameters of Swiss albino mice exposed to fluoride for seven days at daily doses of 190 mg kg^?1 body weight and for 90 days at daily doses of 94 mg kg^?1 body weight is reported. Compared with controls, fluoride exposure decreased erythrocyte (12%–18%), leukocyte (16%) and platelet counts (7%, only for short-term exposure), hemoglobin (2%–17%) and packed cell volume (2%–14%) in Swiss albino mice reared on standard feed but these were altered little in the diet supplement groups. Fluoride exposure decreased lymphocytes in both standard feed (16%–21%) and diet supplement groups (5%–19%, but only after long-term exposure). Fluoride withdrawal led to recovery of the post-treated mice from long-term exposure. Spirulina alone and also in combination with tamarind fruit pulp was found more effective than tamarind alone in reducing hematological disorders in fluoride treated mice.