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Fulltext The diagnosis of scrub typhus: an evaluation

Ramanathan M, Zainal Abidin MN, Balachand V

Med J Malaysia, 1987 Mar;42(1):61-4.
PMID: 3123899

Scrub typhus is an endemic problem in Malaysia. Yet its diagnosis appears to depend heavily on the Wetl-Felix test as the more sophisticated diagnostic procedures are not available routinely. We therefore reviewed our experience with scrub typhus patients treated at the Melaka General Hospital from 1983 to April 1986, to identify those clinical features which are diagnostic of this rickettsial illness. Based on the clinical presentation of our patients and the dramatic response of scrub typhus to Doxycycline, we propose a clinical approach to diagnosis until more specific and cheap diagnostic procedures become available in our laboratories. Otherwise, this rickettsial illness will continue to be under-recognised.

Matched MeSH terms: Antibodies, Bacterial/analysis
Detection of rickettsial antibodies using Weil-Felix (OXK and OX19) antigens and the indirect immunoperoxidase assay

Tay ST, Kamalanathan M, Rohani MY

Southeast Asian J. Trop. Med. Public Health, 2003 Mar;34(1):171-4.
PMID: 12971531
Matched MeSH terms: Antibodies, Bacterial/immunology*
Fulltext Laboratory acquired murine typhus--a case report

Norazah A, Mazlah A, Cheong YM, Kamel AG

Med J Malaysia, 1995 Jun;50(2):177-9.
PMID: 7565191

A 34-year-old laboratory worker developed murine typhus after an accidental splashing of Rickettsia typhi over her right eye and lips. Indirect immunoperoxidase test showed a four-fold increase in titre to Rickettsia typhi. She responded well to doxycycline.

Matched MeSH terms: Antibodies, Bacterial/analysis
Fulltext Serum antibodies to defined carbohydrate antigens during the course of treated leprosy

Gelber RH, Li F, Cho SN, Byrd S, Rajagopalan K, Brennan PJ

Int. J. Lepr. Other Mycobact. Dis., 1989 Dec;57(4):744-51.
PMID: 2681457

Sequential monitoring of 724 sera for antibodies to a neoantigen based on phenolic glycolipid-I (PGL-I) and native lipoarabinomannan (LAM) in 90 leprosy patients undergoing therapy in San Francisco was conducted. Untreated lepromatous patients frequently (91%) had significant antibodies to both moieties. Antibodies were less frequently found in tuberculoid patients (74% to neoantigen and 37% to LAM). In the first 3 years of treatment, average serum antibodies to both moieties fell significantly. Antibodies to LAM fell during each of the first 4 years of therapy, but decreasing antibody levels to the PGL-I neoantigen did not appear to fall consistently after the third year of treatment. A wide variation in the rate of fall of serum antibodies was noted. Sequential changes in the amounts of serum antibodies to the neoantigen and LAM in general paralleled one another but were at times discrepant. Both in San Francisco and Malaysia, skin-smear negative, long-term treated, lepromatous leprosy patients frequently harbored significant antibodies to both PGL-I and LAM.

Matched MeSH terms: Antibodies, Bacterial/analysis*
Development of antibody to Rickettsia tsutsugamushi in soldiers in Malaysia

Brown GW, Shirai A, Groves MG

Trans R Soc Trop Med Hyg, 1983;77(2):225-7.
PMID: 6408770

Malaysian, British and New Zealand soldiers were tested for evidence of infection with Rickettsia tsutsugamushi after several weeks' exposure to the infection during field exercises in Malaysia. 39 (5.0%) of 787 British and New Zealand soldiers developed immunofluorescent antibody (IFA) to R. tsutsugamushi to a titre of 1:50 and two (0.3%) to a titre of 1:100. 11 (1.5%) of 751 Malaysian soldiers also developed low titres less than or equal to 1:100. These low antibody levels were not correlated with clinical disease, and their significance is unknown. Seven (0.9%) of the Malaysians showed an IFA rise to greater than or equal to 1:200, and three of these experienced febrile illnesses, one lasting two weeks. An additional eight Malaysian soldiers had an IFA titre of greater than or equal to 1:400 when first tested and six of these also had a Proteus OXK agglutinin titre of greater than or equal to 1:160, indicating infection shortly before the study.

Matched MeSH terms: Antibodies, Bacterial/biosynthesis*
The longevity of antibody to Rickettsia tsutsugamushi in patients with confirmed scrub typhus

Saunders JP, Brown GW, Shirai A, Huxsoll DL

Trans R Soc Trop Med Hyg, 1980;74(2):253-7.
PMID: 6770503

Serological surveillance for up to two years of 114 patients with laboratory confirmed scrub typhus showed that antibody to Rickettsia tsutsugamushi as demonstrated by the indirect fluorescent antibody test is short-lived. The mean reversion time from mean peak titre (1:499) was 48.9 weeks and the calculated annual reversion rate to a titre less than 1:50 was 61%. This can be used to estimate attack rates based on point prevalence of antibody. The relationship between antibody prevalence and attack rates observed by other workers was confirmed using this model. The possible uses of the finding and its implications in Malaysia are briefly discussed.

Matched MeSH terms: Antibodies, Bacterial/physiology*
A retrospective serological suvey of Royal Marines previously exposed to Pseudomonas pseudomallei in South East Asia

Preston PJ, Lightfoot N, Clarke P

Trans R Soc Trop Med Hyg, 1976;70(4):335-7.
PMID: 1006764

Following the suggestion that it was possible that cases of melioidosis amongst those who had been exposed abroad in the past, might be escaping notice, 487 Royal Marines were examined by indirect haemagglutination studies. Four hundred and eleven of these subjects had served for variable times in areas where melioidosis has been known to occur in Indonesia and Malaya, between 1960 and 1974, occupied in activities in the jungle and paddy fields during which exposure to the disease was to be expected. No evidence of residual subclinical melioidosis was found and it seems unlikely that recrudescent disease will prove to be a problem in the future for English servicemen who have been in South East Asia.

Matched MeSH terms: Antibodies, Bacterial/analysis*
Preliminary study on serum immunoglobulin G responses following intramuscular inoculation of adjuvanted polyhydroxyalkanoate microparticles with Pasteurella multocida vaccine in white rats

Mohamed S, May Amelia TS, Abdullah Amirul AA, Abdul Wahid ME, Bhubalan K

Biologicals, 2021 Jun;71:51-54.
PMID: 33858743 DOI: 10.1016/j.biologicals.2021.03.002

A natural biodegradable polymer, polyhydroxyalkanoate (PHA), was adjuvanted with a vaccine seed to observe the biomaterial's ability in enhancing an immune response in rats. The adjuvant potential of PHA was tested using the whole-killed Pasteurella multocida B:2 (PMB2) vaccine in Sprague Dawley (SD) rats to detect changes in serum immunoglobulin G (IgG) and immunoglobulin M (IgM) responses. A common PHA, poly(3-hydroxybutyrate) [P(3HB)], from Bacillus megaterium UMTKB-1 was constructed into microparticles using the solvent evaporation method. Twelve SD rats were divided into four treatment groups: 1) non-treatment as negative control, 2) P(3HB) adjuvant, 3) PMB2 vaccine, and 4) adjuvanted-P(3HB)/PMB2 vaccine groups, which were intramuscularly vaccinated twice. Immunoglobulins IgG and IgM levels were used as markers of the immune response induced by the adjuvanted-P(3HB)/PMB2 vaccine and analysed over an eight-week study period. The group vaccinated specifically with adjuvanted-P(3HB)/PMB2 vaccine had higher concentrations of immunoglobulins compared to other treatment groups, hence demonstrating the potential of the adjuvant to enhance immune response. Findings showed a need to delay the delivery of the second booster dose to determine the appropriate regime for the adjuvanted-P(3HB)/PMB2 vaccine.

Matched MeSH terms: Antibodies, Bacterial/blood*
Fulltext Leptospirosis among Dengue-Negative Febrile Patients in Selangor, Malaysia

Loong SK, Abd-Majid MA, Teoh BT, Cheh MJ, Khor CS, Chao CC, et al.

Am J Trop Med Hyg, 2022 Aug 17;107(2):397-400.
PMID: 35895409 DOI: 10.4269/ajtmh.20-0656

In recent years, the number of leptospirosis cases, including the number of deaths, has exponentially increased in Malaysia. From June 2016 to February 2018, blood samples of 321 febrile patients with the presumptive diagnosis of dengue-like illness were examined for possible exposure to Leptospira. Two hundred fifty-five blood samples were tested as negative for dengue. Seminested polymerase chain reaction (PCR) and IgM ELISA for leptospirosis were performed. From the samples, an overall prevalence for leptospirosis based on PCR of 4.7% (12/255) was obtained. Eighteen percent (46/255) were positive for anti-Leptospira IgM antibodies. The genome sequences of six of 12 Leptospira PCR-positive samples showed > 97.0% similarity to Leptospira interrogans. One patient's sample consisted of Leptospira and chikungunya virus, suggesting a coinfection. Findings from the study suggest that leptospirosis is prevalent among dengue-negative febrile patients in Malaysia.

Matched MeSH terms: Antibodies, Bacterial
Neutralizing chimeric mouse-human antibodies against Burkholderia pseudomallei protease: expression, purification and characterization

Chan SW, Ong GI, Nathan S

J. Biochem. Mol. Biol., 2004 Sep 30;37(5):556-64.
PMID: 15479619

A recombinant Fab monoclonal antibody (Fab) C37, previously obtained by phage display and biopanning of a random antibody fragment library against Burkholderia pseudomallei protease, was expressed in different strains of Escherichia coli. E. coli strain HB2151 was deemed a more suitable host for Fab expression than other E. coli strains when grown in media supplemented with 0.2 % glycerol. The expressed Fab fragment was purified by affinity chromatography on a Protein G-Sepharose column, and the specificity of the recombinant Fab C37 towards B. pseudomallei protease was proven by Western blotting, enzyme-linked immunosorbent assay (ELISA) and by proteolytic activity neutralization. In addition, polyclonal antibodies against B. pseudomallei protease were produced in rabbits immunized with the protease. These were isolated from high titer serum by affinity chromatography on recombinant-Protein A-Sepharose. Purified polyclonal antibody specificity towards B. pseudomallei protease was proven by Western blotting and ELISA.

Matched MeSH terms: Antibodies, Bacterial/genetics; Antibodies, Bacterial/immunology*; Antibodies, Bacterial/isolation & purification; Antibodies, Bacterial/metabolism
Expression and purification of a recombinant scFv towards the exotoxin of the pathogen, Burkholderia pseudomallei

Lim KP, Li H, Nathan S

J Microbiol, 2004 Jun;42(2):126-32.
PMID: 15357306

A single chain variable fragment (scFv) specific towards B. pseudomallei exotoxin had previously been generated from an existing hybridoma cell line (6E6AF83B) and cloned into the phage display vector pComb3H. In this study, the scFv was subcloned into the pComb3X vector to facilitate the detection and purification of expressed antibodies. Detection was facilitated by the presence of a hemagglutinin (HA) tag, and purification was facilitated by the presence of a histidine tag. The culture was grown at 30 degrees C until log phase was achieved and then induced with 1 mM IPTG in the absence of any additional carbon source. Induction was continued at 30 degrees C for five h. The scFv was discerned by dual processes-direct enzyme-linked immunosorbent assays (ELISA), and Western blotting. When compared to E. coli strains ER2537 and HB2151, scFv expression was observed to be highest in the E. coli strain Top10F'. The expressed scFv protein was purified via nickel-mediated affinity chromatography and results indicated that two proteins a 52 kDa protein, and a 30 kDa protein were co-purified. These antibodies, when blotted against immobilized exotoxin, exhibited significant specificity towards the exotoxin, compared to other B. pseudomallei antigens. Thus, these antibodies should serve as suitable reagents for future affinity purification of the exotoxin.

Matched MeSH terms: Antibodies, Bacterial/analysis; Antibodies, Bacterial/genetics*; Antibodies, Bacterial/immunology*; Antibodies, Bacterial/isolation & purification; Antibodies, Bacterial/chemistry
Fulltext Selection of phage-displayed human antibody fragments specific for CD1b presenting the Mycobacterium tuberculosis glycolipid Ac2SGL

Camacho F, Sarmiento ME, Reyes F, Kim L, Huggett J, Lepore M, et al.

Int J Mycobacteriol, 2016 06;5(2):120-7.
PMID: 27242221 DOI: 10.1016/j.ijmyco.2015.12.002

OBJECTIVE/BACKGROUND: The development of new tools capable of targeting Mycobacterium tuberculosis (Mtb)-infected cells have potential applications in diagnosis, treatment, and prevention of tuberculosis. In Mtb-infected cells, CD1b molecules present Mtb lipids to the immune system (Mtb lipid-CD1b complexes). Because of the lack of CD1b polymorphism, specific Mtb lipid-CD1b complexes could be considered as universal Mtb infection markers. 2-Stearoyl-3-hydroxyphthioceranoyl-2'-sulfate-α-α'-d-trehalose (Ac2SGL) is specific for Mtb, and is not present in other mycobacterial species. The CD1b-Ac2SGL complexes are expressed on the surface of human cells infected with Mtb. The aim of this study was to generate ligands capable of binding these CD1b-Ac2SGL complexes.
METHODS: A synthetic human scFv phage antibody library was used to select phage-displayed antibody fragments that recognized CD1b-Ac2SGL using CD1b-transfected THP-1 cells loaded with Ac2SGL.
RESULTS: One clone, D11-a single, light-variable domain (kappa) antibody (dAbκ11)-showed high relative binding to the Ac2SGL-CD1b complex.
CONCLUSION: A ligand recognizing the Ac2SGL-CD1b complex was obtained, which is a potential candidate to be further tested for diagnostic and therapeutic applications.

Matched MeSH terms: Antibodies, Bacterial/genetics; Antibodies, Bacterial/immunology*
Fulltext Supplementation with natural forms of vitamin E augments antigen-specific TH1-type immune response to tetanus toxoid

Radhakrishnan AK, Mahalingam D, Selvaduray KR, Nesaretnam K

Biomed Res Int, 2013;2013:782067.
PMID: 23936847 DOI: 10.1155/2013/782067

This study compared the ability of three forms of vitamin E [tocotrienol-rich fraction (TRF), alpha-tocopherol (α-T), and delta-tocotrienol (δ-T3)] to enhance immune response to tetanus toxoid (TT) immunisation in a mouse model. Twenty BALB/c mice were divided into four groups of five mice each. The mice were fed with the different forms of vitamin E (1 mg) or vehicle daily for two weeks before they were given the TT vaccine [4 Lf] intramuscularly (i.m.). Booster vaccinations were given on days 28 and 42. Serum was collected (days 0, 28, and 56) to quantify anti-TT levels. At autopsy, splenocytes harvested were cultured with TT or mitogens. The production of anti-TT antibodies was augmented (P < 0.05) in mice that were fed with δ-T3 or TRF compared to controls. The production of IFN-γ and IL-4 by splenocytes from the vitamin E treated mice was significantly (P < 0.05) higher than that from controls. The IFN-γ production was the highest in animals supplemented with δ-T3 followed by TRF and finally α-T. Production of TNF-α was suppressed in the vitamin E treated group compared to vehicle-supplemented controls. Supplementation with δ-T3 or TRF can enhance immune response to TT immunisation and production of cytokines that promote cell-mediated (TH1) immune response.

Matched MeSH terms: Antibodies, Bacterial/biosynthesis; Antibodies, Bacterial/immunology*
Fulltext Seroprevalance of brucellosis among suspected cases in Malaysia

Jama'ayah MZ, Heu JY, Norazah A

Malays J Pathol, 2011 Jun;33(1):31-4.
PMID: 21874749 MyJurnal

Brucellosis is a zoonotic disease which can be transmitted by direct or indirect contact with infected animal or their products. It is an important public health problem but little is known on brucellosis in the Malaysian population. The aim of this study was to determine the presence of Brucella antibodies using commercial Brucella IgG and IgM ELISA kits (Vircell, SL, Barcelona Spain). A total of 184 sera from suspected patients were received from 16 hospitals in Malaysia over the years 2004 to 2009. Only 10 serum samples (5.4%) were positive for Brucella antibodies in which 5 showed the presence of both IgM and IgG. Most of the positive patients were occupationally involved with animals. This study suggests the seroprevalance of brucellosis among individuals who have contact with infected animals in Malaysia is low.

Matched MeSH terms: Antibodies, Bacterial/analysis; Antibodies, Bacterial/blood*
Antibody response to Helicobacter pylori excretory antigen and the cross reaction study

Sasidharan S, Uyub AM

J Immunoassay Immunochem, 2009;30(1):70-81.
PMID: 19117203 DOI: 10.1080/15321810802569477

Helicobacter pylori is recognized as a major case of gastritis and peptic ulcer and a key factor in the development of gastric cancer, gastric lymphoma, and non-ulcerative dyspepsia in man. The detection of antibodies specific for strains of H. pylori has demonstrated the value of serology for providing evidence of infection. The present study was conducted to detect the antigenic proteins of excretory antigen of H. pylori with Western blotting and examine whether anti-H. pylori IgG and IgA antibodies from H. pylori positive patients cross-react with antigens from other common bacterial pathogens. By using SDS-PAGE, 20 different proteins were found in the excretory antigen. By Western blotting and absorption studies, there were indications that anti-H. pylori IgA antibodies directed against 54 kDa, 50 kDa and 27 kDa cross-reacted with antigens from other bacteria, and that H. pylori proteins of 99 kDa, 88 kDa and 81 kDa possibly shared similar epitope with antigens of other pathogens not tested in the absorption studies. The cross-reactivity occurred in this study was not significantly affect the performance of the in-house ELISA.

Matched MeSH terms: Antibodies, Bacterial/blood; Antibodies, Bacterial/immunology*
Relationship between active protection in vaccinated buffaloes against haemorrhagic septicaemia and passive mouse protection test or serum antibody titres

Chandrasekaran S, Kennett L, Yeap PC, Muniandy N, Rani B, Mukkur TK

Vet Microbiol, 1994 Aug 15;41(4):303-9.
PMID: 7801530

The relationship between the standard passive mouse protection test or serum antibody titres measured by indirect haemagglutination or enzyme-linked immunosorbent assays and active protection in buffaloes immunized with different types of haemorrhagic septicaemia bacterins was investigated. Groups of 2-3 buffaloes were immunized with the bacterins currently in use in Asia, viz., broth bacterin (BB), alum precipitated vaccine (APV) and oil adjuvant vaccine (OAV) either subcutaneously (BB, APV) or intramuscularly (OAV) and challenged subcutaneously with virulent organisms at different periods post-immunization. Although the passive mouse protection and indirect haemagglutination tests carried out with the pre-challenge sera from vaccinated buffaloes revealed no relationship with active protection in buffaloes, a relationship was observed between the ELISA antibody titres and protection. In contrast, a dose-response relationship was observed between the homologous active and passive mouse protection test.

Matched MeSH terms: Antibodies, Bacterial/biosynthesis; Antibodies, Bacterial/blood
Fluorescence-based immunoassay for the detection of Xanthomonas oryzae pv. oryzae in rice leaf

Awaludin N, Abdullah J, Salam F, Ramachandran K, Yusof NA, Wasoh H

Anal Biochem, 2020 12 01;610:113876.
PMID: 32750357 DOI: 10.1016/j.ab.2020.113876

The identification of rice bacterial leaf blight disease requires a simple, rapid, highly sensitive, and quantitative approach that can be applied as an early detection monitoring tool in rice health. This paper highlights the development of a turn-off fluorescence-based immunoassay for the early detection of Xanthomonas oryzae pv. oryzae (Xoo), a gram-negative bacterium that causes rice bacterial leaf blight disease. Antibodies against Xoo bacterial cells were produced as specific bio-recognition molecules and the conjugation of these antibodies with graphene quantum dots and gold nanoparticles was performed and characterized, respectively. The combination of both these bio-probes as a fluorescent donor and metal quencher led to changes in the fluorescence signal. The immunoreaction between AntiXoo-GQDs, Xoo cells, and AntiXoo-AuNPs in the immuno-aggregation complex led to the energy transfer in the turn-off fluorescence-based quenching system. The change in fluorescence intensity was proportional to the logarithm of Xoo cells in the range of 100-105 CFU mL-1. The limit of detection was achieved at 22 CFU mL-1 and the specificity test against other plant disease pathogens showed high specificity towards Xoo. The detection of Xoo in real plant samples was also performed in this study and demonstrated satisfactory results.

Matched MeSH terms: Antibodies, Bacterial/immunology; Antibodies, Bacterial/chemistry
Fulltext Prevalence of anti-Leptospira antibodies and associated risk factors in the Malaysian refugee communities

Mohd Hanapi IR, Sahimin N, Maackara MJB, Annisa AS, Abdul Mutalib RNS, Lewis JW, et al.

BMC Infect Dis, 2021 Nov 01;21(1):1128.
PMID: 34724919 DOI: 10.1186/s12879-021-06830-0

BACKGROUND: Refugees in Malaysia, who are afflicted by poverty, conflict and poor health, are vulnerable to a range of zoonotic infections in the deprived environmental and social conditions under which they live. Exposure to infections such as leptospirosis, for which rodents are primary hosts, is of particular concern.
METHODS: A wellness program was conducted to determine the presence of antibodies against Leptospira (seroprevalence) in 11 refugee community schools and centers in the Klang Valley, Malaysia. A total of 433 samples were assessed for IgG and IgM antibodies against Leptospira, using enzyme-linked immunosorbent assays (ELISA).
RESULTS: Overall Leptospira seroprevalence was 24.7%, with 3.0% being seropositive for anti-Leptospira IgG and 21.7% for anti-Leptospira IgM. Factors significantly associated with overall Leptospira seroprevalence included: age, ethnicity, pet ownership, knowledge of disease and awareness of disease fatality. For IgM seroprevalence, significant risk factors included sex, ethnicity, eating habits with hands, pet ownership, the presence of rats, walking in bare feet and water recreation visits.
CONCLUSIONS: These findings highlight the need for improvements in health and well-being among the refugee community through disease awareness programs and provision of healthy behavior programs, particularly in hygiene and sanitation through community engagement activities.

Matched MeSH terms: Antibodies, Bacterial
Serum antibody response against periodontal bacteria and coronary heart disease: Systematic review and meta-analysis

Joshi C, Bapat R, Anderson W, Dawson D, Cherukara G, Hijazi K

J Clin Periodontol, 2021 12;48(12):1570-1586.
PMID: 34545592 DOI: 10.1111/jcpe.13550

AIM: The present systematic review and meta-analysis assessed the strength of a reported association between elevated serum anti-periodontal bacterial antibody responses and coronary heart disease (CHD).
MATERIALS AND METHODS: Twenty original studies were identified after systematically searching five databases. The majority (n = 11) compared serum anti-Porphyromonas gingivalis (Pg) and/or anti-Aggregatibacter actinomycetemcomitans (Aa) IgG antibody responses between CHD patients and control participants. The strength of the association between serum anti-Pg antibodies and CHD (n = 10) and serum anti-Aa antibodies and CHD (n = 6) was investigated using a meta-analysis approach separately.
RESULTS: Most studies (61%) reported that the serum IgG antibody responses were elevated in CHD patients than in controls. The meta-analyses showed a significant association between elevated serum IgG antibody responses (anti-Pg and anti-Aa) and CHD, with pooled odds ratios of 1.23 [95% confidence interval (CI): 1.09-1.38, p = .001] and 1.25 (95% CI: 1.04-1.47, p = .0004), respectively.
CONCLUSIONS: A modest increase of CHD events in individuals with higher serum anti-Pg and anti-Aa IgG antibody responses may support their use as potential biomarkers to detect and monitor at-risk populations. However, the observed inconsistencies with the design and interpretation of immunoassays warrant standardization of the immunoassays assessing antibody responses against periodontal bacteria.

Matched MeSH terms: Antibodies, Bacterial
Fulltext In-house ELISA screening using a locally-isolated Leptospira in Malaysia: determination of its cut-off points

Tan XT, Amran F, Chee Cheong K, Ahmad N

BMC Infect Dis, 2014;14:563.
PMID: 25338815 DOI: 10.1186/s12879-014-0563-7

Leptospirosis is a zoonotic disease caused by Leptospira species and is distributed globally. Microscopic agglutination test (MAT) is the serological 'gold standard' for diagnosis of leptospirosis but it is time-consuming and labour-intensive. An alternative serological method that is rapid, sensitive and specific is important for early treatment to reduce morbidity and mortality. The use of local Leptospira isolation may improve the sensitivity and specificity of the test because it may varies from one geographical region to another region. The objective of this study was to determine the sensitivity, specificity and cut-off points for an in-house Immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) using a locally isolated Leptospiral strain IMR/175 as the antigen for the detection of anti-Leptospiral IgM.

Matched MeSH terms: Antibodies, Bacterial/blood

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